DNase variants

ABSTRACT

The present invention relates to polypeptide variants and methods for obtaining variants. The present invention also relates to polynucleotides encoding the variants; nucleic acid constructs, vectors, and host cells comprising the polynucleotides; and methods of using the variants.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No. 16/571,907filed on Sep. 16, 2019, now abandoned, which is a divisional of U.S.application Ser. No. 15/737,184 filed on Dec. 15, 2017, now U.S. Pat.No. 10,479,981, which is a 35 U.S.C. 371 national application ofinternational application no. PCT/EP2016/074747 filed Oct. 14, 2016,which claims priority or the benefit under 35 U.S.C. 119 of Danishapplication no. PA 2015 00632 filed Oct. 14, 2015, the contents of whichare fully incorporated herein by reference.

REFERENCE TO A SEQUENCE LISTING

This application contains a Sequence Listing in computer readable form,which is incorporated herein by reference.

BACKGROUND OF THE INVENTION Field of the Invention

The present invention relates to novel DNase variants exhibitingalterations relative to the parent DNase in one or more propertiesincluding: wash performance, detergent stability and/or storagestability. The variants of the invention are suitable for use incleaning processes and detergent compositions, such as laundrycompositions and dish wash compositions, including hand wash andautomatic dish wash compositions. The present invention also relates toisolated DNA sequences encoding the variants, expression vectors, hostcells, and methods for producing and using the DNase variants of theinvention.

Description of the Related Art

Microorganisms generally live attached to surfaces in many natural,industrial, and medical environments, encapsulated by extracellularsubstances including biopolymers and macromolecules. The resulting layerof slime encapsulated microorganism is termed a biofilm. Biofilms arethe predominant mode of growth of bacteria in the natural environment,and bacteria growing in biofilms exhibit distinct physiologicalproperties. Compared to their planktonically grown counterparts, thebacteria in a biofilm are more resistant to antibiotics, UV irradiation,detergents and the host immune response.

A biofilm may include one or more microorganisms, includinggram-positive and gram-negative bacteria, algae, protozoa, and/or yeastor filamentous fungi and viruses and/or bacteriophage. Examples ofproblematic biofilms are dental plaque, infections on medical implants,but also the initial fouling on ship hulls. Biofilms are attributed tothe pathogenesis of many infections in humans and are a significantproblem in industry in terms of biofouling of exposed surfaces, wherebiofilm colonisation can form the base component of a localisedecosystem which can disrupt and interfere with industrial processes andcomponents.

When laundry items like T-shirts or sportswear are used, they areexposed to bacteria from the body of the user and from the rest of theenvironment in which they are used. Some of these bacteria are capableof adhering to the laundry item and form a biofilm on the item. Thepresence of bacteria implies that the laundry items become sticky andtherefore soil adheres to the sticky areas. This soil has showndifficult to remove by commercially available detergent compositions.Further, when very dirty laundry items are washed together with lessdirty laundry items the dirt present in the wash liquor tend to stick tothe biofilm. As a result hereof the laundry item is more “soiled” afterwash than before wash. Further, these bacteria are a source of bad odor,which develops after use of the laundry item. The bad odor is difficultto remove and may remain even after wash. The reason for this bad odoris adhesion of bacteria to the textile surface. Because of the adhesionto the textile, the bacteria may remain even after wash, and continue tobe a source of bad odor.

International patent application WO2011/098579 (Newcastle UNIV.) andWO2014/087011 (Novozymes A/S) concern bacterial deoxyribonucleasecompounds and methods for biofilm disruption and prevention.

SUMMARY OF THE INVENTION

The present invention relates to a DNase variant having at least oneimproved property compared to SEQ ID NO: 1, comprising a modification atone or more positions selected from the list consisting of 1, 2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23,24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41,42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 53, 54, 55, 56, 57, 58, 59, 60,61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111,112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125,126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139,140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153,154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167,168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181,182, 183, 185, 186, 187, 189, 190, 191, 192, 193, 194, 195, 196, 197,198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211,212, 213, 214, 215, 216, 217, 218, 219, 220 and 221, wherein the varianthas an amino acid sequence which is at least 60%, at least 70%, at least80%, at least 85%, at least 90% or at least 95% identical to SEQ ID NO:1.

The invention further relates to variants of a DNase parent with atleast 60% identity to SEQ ID NO: 1 wherein said variant comprises atleast one of the following modifications compared to SEQ ID NO: 1: V1*,V1A, V1D, V1E, V1F, V1G, V1H, V1I, V1K, V1L, V1M, V1N, V1P, V1Q, V1R,V1S, V1T, V1W, V1Y, P2*, P2A, P2D, P2E, P2F, P2G, P2H, P2I, P2K, P2L,P2M, P2N, P2Q, P2R, P2S, P2T, P2V, P2W, P2Y, V3*, V3A, V3C, V3D, V3E,V3F, V3G, V3H, V3I, V3K, V3L, V3M, V3N, V3P, V3R, V3S, V3T, V3W, V3Y,N4*, N4A, N4D, N4E, N4F, N4G, N4H, N4I, N4K, N4L, N4M, N4P, N4Q, N4R,N4S, N4T, N4V, N4W, N4Y, P5*, P5A, P5D, P5E, P5F, P5G, P5H, P5I, P5K,P5L, P5M, P5N, P5Q, P5R, P5S, P5T, P5V, P5W, P5Y, E6*, E6A, E6D, E6F,E6G, E6H, E6I, E6K, E6L, E6M, E6N, E6P, E6Q, E6R, E6S, E6T, E6V, E6W,E6Y, P7*, P7A, P7D, P7E, P7F, P7G, P7H, P7I, P7K, P7L, P7M, P7N, P7Q,P7R, P7S, P7T, P7V, P7W, P7Y, D8*, D8A, D8E, D8F, D8G, D8H, D8I, D8K,D8L, D8M, D8N, D8P, D8Q, D8R, D8S, D8T, D8V, D8W, D8Y, A9*, A9D, A9E,A9F, A9G, A9H, A9I, A9K, A9L, A9M, A9N, A9P, A9Q, A9R, A9S, A9T, A9V,A9W, A9Y, T10*, T10A, T10D, T10E, T10F, T10G, T10H, T10I, T10K, T10L,T10M, T10N, T10P, T10Q, T10R, T10S, T10V, T10W, T10Y, S11*, S11A, S11D,S11E, S11F, S11G, S11H, S11I, S11K, S11L, S11M, S11N, S11P, S11Q, S11R,S11T, S11V, S11W, S11Y, V12*, V12A, V12D, V12E, V12F, V12G, V12H, V12I,V12K, V12L, V12M, V12N, V12P, V12Q, V12R, V12S, V12T, V12W, V12Y, E13*,E13A, E13D, E13F, E13G, E13H, E13I, E13K, E13L, E13M, E13N, E13P, E13Q,E13R, E13S, E13T, E13V, E13W, E13Y, N14*, N14A, N14D, N14E, N14F, N14G,N14H, N14I, N14K, N14L, N14M, N14P, N14Q, N14R, N14S, N14T, N14V, N14W,N14Y, V15*, V15A, V15D, V15E, V15F, V15G, V15H, V15I, V15K, V15L, V15M,V15N, V15P, V15Q, V15R, V15S, V15T, V15W, V15Y, A16*, A16D, A16E, A16F,A16G, A16H, A16I, A16K, A16L, A16M, A16N, A16P, A16Q, A16R, A16S, A16T,A16V, A16W, A16Y, L17*, L17A, L17D, L17E, L17F, L17G, L17H, L17I, L17K,L17M, L17N, L17P, L17Q, L17R, L17S, L17T, L17V, L17W, L17Y, K18*, K18A,K18D, K18E, K18F, K18G, K18H, K18I, K18L, K18M, K18N, K18P, K18Q, K18R,K18S, K18T, K18V, K18W, K18Y, T19*, T19A, T19D, T19E, T19F, T19G, T19H,T19I, T19K, T19L, T19M, T19N, T19P, T19Q, T19R, T19S, T19V, T19W, T19Y,G20*, G20A, G20D, G20E, G20F, G20H, G20I, G20K, G20L, G20M, G20N, G20P,G20Q, G20R, G20S, G20T, G20V, G20W, G20Y, S21*, S21A, S21D, S21E, S21F,S21G, S21H, S21I, S21K, S21L, S21M, S21N, S21P, S21Q, S21R, S21T, S21V,S21W, S21Y, G22*, G22A, G22D, G22E, G22F, G22H, G22I, G22K, G22L, G22M,G22N, G22P, G22Q, G22R, G22S, G22T, G22V, G22W, G22Y, D23*, D23A, D23E,D23F, D23G, D23H, D23I, D23K, D23L, D23M, D23N, D23P, D23Q, D23R, D23S,D23T, D23V, D23W, D23Y, S24*, S24A, S24D, S24E, S24F, S24G, S24H, S24I,S24K, S24L, S24M, S24N, S24P, S24Q, S24R, S24T, S24V, S24W, S24Y, Q25*,Q25A, Q25D, Q25E, Q25F, Q25G, Q25H, Q25I, Q25K, Q25L, Q25M, Q25N, Q25P,Q25R, Q25S, Q25T, Q25V, Q25W, Q25Y, S26*, S26A, S26D, S26E, S26F, S26G,S26H, S26I, S26K, S26L, S26M, S26N, S26P, S26Q, S26R, S26T, S26V, S26W,S26Y, D27*, D27A, D27E, D27F, D27G, D27H, D27I, D27K, D27L, D27M, D27N,D27P, D27Q, D27R, D27S, D27T, D27V, D27W, D27Y, P28*, P28A, P28D, P28E,P28F, P28G, P28H, P28I, P28K, P28L, P28M, P28N, P28Q, P28R, P28S, P28T,P28V, P28W, P28Y, I29*, I29A, I29D, I29E, I29F, I29G, I29H, I29K, I29L,I29M, I29N, I29P, I29Q, I29R, I29S, I29T, I29V, I29W, I29Y, K30*, K30A,K30D, K30E, K30F, K30G, K30H, K30I, K30L, K30M, K30N, K30P, K30Q, K30R,K30S, K30T, K30V, K30W, K30Y, A31*, A31D, A31E, A31F, A31G, A31H, A31I,A31K, A31L, A31M, A31N, A31P, A31Q, A31R, A31S, A31T, A31V, A31W, A31Y,D32*, D32A, D32E, D32F, D32G, D32H, D32I, D32K, D32L, D32M, D32N, D32P,D32Q, D32R, D32S, D32T, D32V, D32W, D32Y, L33*, L33A, L33D, L33E, L33F,L33G, L33H, L33I, L33K, L33M, L33N, L33P, L33Q, L33R, L33S, L33T, L33V,L33W, L33Y, E34*, E34A, E34D, E34F, E34G, E34H, E34I, E34K, E34L, E34M,E34N, E34P, E34Q, E34R, E34S, E34T, E34V, E34W, E34Y, V35*, V35A, V35D,V35E, V35F, V35G, V35H, V35I, V35K, V35L, V35M, V35N, V35P, V35Q, V35R,V35S, V35T, V35W, V35Y, K36*, K36A, K36D, K36E, K36F, K36G, K36H, K36I,K36L, K36M, K36N, K36P, K36Q, K36R, K36S, K36T, K36V, K36W, K36Y, G37*,G37A, G37D, G37E, G37F, G37H, G37I, G37K, G37L, G37M, G37N, G37P, G37Q,G37R, G37S, G37T, G37V, G37W, G37Y, Q38*, Q38A, Q38D, Q38E, Q38F, Q38G,Q38H, Q38I, Q38K, Q38L, Q38M, Q38N, Q38P, Q38R, Q38S, Q38T, Q38V, Q38W,Q38Y, S39*, S39A, S39D, S39E, S39F, S39G, S39H, S39I, S39K, S39L, S39M,S39N, S39P, S39Q, S39R, S39T, S39V, S39W, S39Y, A40*, A40D, A40E, A40F,A40G, A40H, A40I, A40K, A40L, A40M, A40N, A40P, A40Q, A40R, A40S, A40T,A40V, A40W, A40Y, L41*, L41A, L41D, L41E, L41F, L41G, L41H, L41I, L41K,L41M, L41N, L41P, L41Q, L41R, L41S, L41T, L41V, L41W, L41Y, P42*, P42A,P42D, P42E, P42F, P42G, P42H, P42I, P42K, P42L, P42M, P42N, P42Q, P42R,P42S, P42T, P42V, P42W, P42Y, F43*, F43A, F43D, F43E, F43G, F43H, F43I,F43K, F43L, F43M, F43N, F43P, F43Q, F43R, F43S, F43T, F43V, F43W, F43Y,D44*, D44A, D44E, D44F, D44G, D44H, D44I, D44K, D44L, D44M, D44N, D44P,D44Q, D44R, D44S, D44T, D44V, D44W, D44Y, V45*, V45A, V45D, V45E, V45F,V45G, V45H, V45I, V45K, V45L, V45M, V45N, V45P, V45Q, V45R, V45S, V45T,V45W, V45Y, D46*, D46A, D46E, D46F, D46G, D46H, D46I, D46K, D46L, D46M,D46N, D46P, D46Q, D46R, D46S, D46T, D46V, D46W, D46Y, C47A, C47D, C47E,C47F, C47G, C47H, C47I, C47K, C47L, C47M, C47N, C47P, C47Q, C47R, C47S,C47T, C47V, C47W, C47Y, W48*, W48A, W48D, W48E, W48F, W48G, W48H, W48I,W48K, W48L, W48M, W48N, W48P, W48Q, W48R, W48S, W48T, W48V, W48Y, A49*,A49D, A49E, A49F, A49G, A49H, A49I, A49K, A49L, A49M, A49N, A49P, A49Q,A49R, A49S, A49T, A49V, A49W, A49Y, 150*, 150A, I50D, I50E, I50F, I50G,I50H, I50K, I50L, I50M, I50N, I50P, I50Q, I50R, I50S, I50T, I50V, I50W,150Y, L51*, L51A, L51D, L51E, L51F, L51G, L51H, L51I, L51K, L51M, L51N,L51P, L51Q, L51R, L51S, L51T, L51V, L51W, L51Y, K53*, K53A, K53D, K53E,K53F, K53G, K53H, K53I, K53L, K53M, K53N, K53P, K53Q, K53R, K53S, K53T,K53V, K53W, K53Y, G54*, G54A, G54D, G54E, G54F, G54H, G54I, G54K, G54L,G54M, G54N, G54P, G54Q, G54R, G54S, G54T, G54V, G54W, G54Y, A55*, A55D,A55E, A55F, A55G, A55H, A55I, A55K, A55L, A55M, A55N, A55P, A55Q, A55R,A55S, A55T, A55V, A55W, A55Y, P56*, P56A, P56D, P56E, P56F, P56G, P56H,P56I, P56K, P56L, P56M, P56N, P56Q, P56R, P56S, P56T, P56V, P56W, P56Y,N57*, N57A, N57D, N57E, N57F, N57G, N57H, N57I, N57K, N57L, N57M, N57P,N57Q, N57R, N57S, N57T, N57V, N57W, N57Y, V58*, V58A, V58D, V58E, V58F,V58G, V58H, V58I, V58K, V58L, V58M, V58N, V58P, V58Q, V58R, V58S, V58T,V58W, V58YL59*, L59A, L59D, L59E, L59F, L59G, L59H, L59I, L59K, L59M,L59N, L59P, L59Q, L59R, L59S, L59T, L59V, L59W, L59Y, Q60*, Q60A, Q60D,Q60E, Q60F, Q60G, Q60H, Q60I, Q60K, Q60L, Q60M, Q60N, Q60P, Q60R, Q60S,Q60T, Q60V, Q60W, Q60Y, R61*, R61A, R61D, R61E, R61F, R61G, R61H, R61I,R61K, R61L, R61M, R61N, R61P, R61Q, R61S, R61T, R61V, R61W, R61Y, V62*,V62A, V62D, V62E, V62F, V62G, V62H, V62I, V62K, V62L, V62M, V62N, V62P,V62Q, V62R, V62S, V62T, V62W, V62Y, N63*, N63A, N63D, N63E, N63F, N63G,N63H, N63I, N63K, N63L, N63M, N63P, N63Q, N63R, N63S, N63T, N63V, N63W,N63Y, E64*, E64A, E64D, E64F, E64G, E64H, E64I, E64K, E64L, E64M, E64N,E64P, E64Q, E64R, E64S, E64T, E64V, E64W, E64Y, K65*, K65A, K65D, K65E,K65F, K65G, K65H, K65I, K65L, K65M, K65N, K65P, K65Q, K65R, K65S, K65T,K65V, K65W, K65Y, T66*, T66A, T66D, T66E, T66F, T66G, T66H, T66I, T66K,T66L, T66M, T66N, T66P, T66Q, T66R, T66S, T66V, T66W, T66Y, K67*, K67A,K67D, K67E, K67F, K67G, K67H, K67I, K67L, K67M, K67N, K67P, K67Q, K67R,K67S, K67T, K67V, K67W, K67Y, N68*, N68A, N68D, N68E, N68F, N68G, N68H,N68I, N68K, N68L, N68M, N68P, N68Q, N68R, N68S, N68T, N68V, N68W, N68Y,S69*, S69A, S69D, S69E, S69F, S69G, S69H, S69I, S69K, S69L, S69M, S69N,S69P, S69Q, S69R, S69T, S69V, S69W, S69Y, N70*, N70A, N70D, N70E, N70F,N70G, N70H, N70I, N70K, N70L, N70M, N70P, N70Q, N70R, N70S, N70T, N70V,N70W, N70Y, R71*, R71A, R71D, R71E, R71F, R71G, R71H, R71I, R71K, R71L,R71M, R71N, R71P, R71Q, R71S, R71T, R71V, R71W, R71Y, D72*, D72A, D72E,D72F, D72G, D72H, D72I, D72K, D72L, D72M, D72N, D72P, D72Q, D72R, D72S,D72T, D72V, D72W, D72Y, R73*, R73A, R73D, R73E, R73F, R73G, R73H, R73I,R73K, R73L, R73M, R73N, R73P, R73Q, R73S, R73T, R73V, R73W, R73Y, S74*,S74A, S74D, S74E, S74F, S74G, S74H, S74I, S74K, S74L, S74M, S74N, S74P,S74Q, S74R, S74T, S74V, S74W, S74Y, G75*, G75A, G75D, G75E, G75F, G75H,G75I, G75K, G75L, G75M, G75N, G75P, G75Q, G75R, G75S, G75T, G75V, G75W,G75Y, A76*, A76D, A76E, A76F, A76G, A76H, A76I, A76K, A76L, A76M, A76N,A76P, A76Q, A76R, A76S, A76T, A76V, A76W, A76Y, N77*, N77A, N77D, N77E,N77F, N77G, N77H, N77I, N77K, N77L, N77M, N77P, N77Q, N77R, N77S, N77T,N77V, N77W, N77Y, K78*, K78A, K78D, K78E, K78F, K78G, K78H, K78I, K78L,K78M, K78N, K78P, K78Q, K78R, K78S, K78T, K78V, K78W, K78Y, G79*, G79A,G79D, G79E, G79F, G79H, G79I, G79K, G79L, G79M, G79N, G79P, G79Q, G79R,G79S, G79T, G79V, G79W, G79Y, P80*, P80A, P80D, P80E, P80F, P80G, P80H,P80I, P80K, P80L, P80M, P80N, P80Q, P80R, P80S, P80T, P80V, P80W, P80Y,F81*, F81A, F81D, F81E, F81G, F81H, F81I, F81K, F81L, F81M, F81N, F81P,F81Q, F81R, F81S, F81T, F81V, F81W, F81Y, K82*, K82A, K82D, K82E, K82F,K82G, K82H, K82I, K82L, K82M, K82N, K82P, K82Q, K82R, K82S, K82T, K82V,K82W, K82Y, D83*, D83A, D83E, D83F, D83G, D83H, D83I, D83K, D83L, D83M,D83N, D83P, D83Q, D83R, D83S, D83T, D83V, D83W, D83Y, P84*, P84A, P84D,P84E, P84F, P84G, P84H, P84I, P84K, P84L, P84M, P84N, P84Q, P84R, P84S,P84T, P84V, P84W, P84Y, Q85*, Q85A, Q85D, Q85E, Q85F, Q85G, Q85H, Q85I,Q85K, Q85L, Q85M, Q85N, Q85P, Q85R, Q85S, Q85T, Q85V, Q85W, Q85Y, K86*,K86A, K86D, K86E, K86F, K86G, K86H, K86I, K86L, K86M, K86N, K86P, K86Q,K86R, K86S, K86T, K86V, K86W, K86Y, W87*, W87A, W87D, W87E, W87F, W87G,W87H, W87I, W87K, W87L, W87M, W87N, W87P, W87Q, W87R, W87S, W87T, W87V,W87Y, G88*, G88A, G88D, G88E, G88F, G88H, G88I, G88K, G88L, G88M, G88N,G88P, G88Q, G88R, G88S, G88T, G88V, G88W, G88Y, I89*, 189A, I89D, I89E,I89F, I89G, I89H, I89K, I89L, I89M, I89N, I89P, I89Q, I89R, I89S, I89T,I89V, I89W, I89Y, K90*, K90A, K90D, K90E, K90F, K90G, K90H, K90I, K90L,K90M, K90N, K90P, K90Q, K90R, K90S, K90T, K90V, K90W, K90Y, A91*, A91D,A91E, A9I, F, A91G, A91H, A91I, A91K, A91L, A91M, A91N, A91P, A91Q,A91R, A91S, A91T, A91V, A91W, A91Y, L92*, L92A, L92D, L92E, L92F, L92G,L92H, L92I, L92K, L92M, L92N, L92P, L92Q, L92R, L92S, L92T, L92V, L92W,L92Y, P93*, P93A, P93D, P93E, P93F, P93G, P93H, P93I, P93K, P93L, P93M,P93N, P93Q, P93R, P93S, P93T, P93V, P93W, P93Y, P94*, P94A, P94D, P94E,P94F, P94G, P94H, P94I, P94K, P94L, P94M, P94N, P94Q, P94R, P94S, P94T,P94V, P94W, P94Y, K95*, K95A, K95D, K95E, K95F, K95G, K95H, K95I, K95L,K95M, K95N, K95P, K95Q, K95R, K95S, K95T, K95V, K95W, K95Y, N96*, N96A,N96D, N96E, N96F, N96G, N96H, N96I, N96K, N96L, N96M, N96P, N96Q, N96R,N96S, N96T, N96V, N96W, N96Y, P97*, P97A, P97D, P97E, P97F, P97G, P97H,P97I, P97K, P97L, P97M, P97N, P97Q, P97R, P97S, P97T, P97V, P97W, P97Y,S98*, S98A, S98D, S98E, S98F, S98G, S98H, S98I, S98K, S98L, S98M, S98N,S98P, S98Q, S98R, S98T, S98V, S98W, S98Y, W99*, W99A, W99D, W99E, W99F,W99G, W99H, W99I, W99K, W99L, W99M, W99N, W99P, W99Q, W99R, W99S, W99T,W99V, W99Y, S100*, S100A, S100D, S100E, S100F, S100G, S100H, S100I,S100K, S100L, S100M, S100N, S100P, S100Q, S100R, S100T, S100V, S100W,S100Y, A101*, A101D, A101E, A101F, A101G, A101H, A101I, A101K, A101L,A101M, A101N, A101P, A101Q, A101R, A101S, A101T, A101V, A101W, A101Y,Q102*, Q102A, Q102D, Q102E, Q102F, Q102G, Q102H, Q102I, Q102K, Q102L,Q102M, Q102N, Q102P, Q102R, Q102S, Q102T, Q102V, Q102W, Q102Y, D103*,D103A, D103E, D103F, D103G, D103H, D103I, D103K, D103L, D103M, D103N,D103P, D103Q, D103R, D103S, D103T, D103V, D103W, D103Y, F104*, F104A,F104D, F104E, F104G, F104H, F104I, F104K, F104L, F104M, F104N, F104P,F104Q, F104R, F104S, F104T, F104V, F104W, F104Y, K105*, K105A, K105D,K105E, K105F, K105G, K105H, K105I, K105L, K105M, K105N, K105P, K105Q,K105R, K105S, K105T, K105V, K105W, K105Y, S106*, S106A, S106D, S106E,S106F, S106G, S106H, S106I, S106K, S106L, S106M, S106N, S106P, S106Q,S106R, S106T, S106V, S106W, S106Y, P107*, P107A, P107D, P107E, P107F,P107G, P107H, P107I, P107K, P107L, P107M, P107N, P107Q, P107R, P107S,P107T, P107V, P107W, P107Y, E108*, E108A, E108D, E108F, E108G, E108H,E108I, E108K, E108L, E108M, E108N, E108P, E108Q, E108R, E108S, E108T,E108V, E108W, E108Y, E109*, E109A, E109D, E109F, E109G, E109H, E109I,E109K, E109L, E109M, E109N, E109P, E109Q, E109R, E109S, E109T, E109V,E109W, E109Y, Y110*, Y110A, Y110D, Y110E, Y110F, Y110G, Y110H, Y110I,Y110K, Y110L, Y110M, Y110N, Y110P, Y110Q, Y110R, Y110S, Y110T, Y110V,Y110W, A111*, A111D, A111E, A111F, A111G, A111H, A111I, A111K, A111L,A111M, A111N, A111P, A111Q, A111R, A111S, A111T, A111V, A111W, A111Y,F112*, F112A, F112D, F112E, F112G, F112H, F112I, F112K, F112L, F112M,F112N, F112P, F112Q, F112R, F112S, F112T, F112V, F112W, F112Y, A113*,A113D, A113E, A113F, A113G, A113H, A113I, A113K, A113L, A113M, A113N,A113P, A113Q, A113R, A113S, A113T, A113V, A113W, A113Y, S114*, S114A,S114D, S114E, S114F, S114G, S114H, S114I, S114K, S114L, S114M, S114N,S114P, S114Q, S114R, S114T, S114V, S114W, S114Y, S115*, S115A, S115D,S115E, S115F, S115G, S115H, S115I, S115K, S115L, S115M, S115N, S115P,S115Q, S115R, S115T, S115V, S115W, S115Y, L116*, L116A, L116D, L116E,L116F, L116G, L116H, L116I, L116K, L116M, L116N, L116P, L116Q, L116R,L116S, L116T, L116V, L116W, L116Y, Q117*, Q117A, Q117D, Q117E, Q117F,Q117G, Q117H, Q117I, Q117K, Q117L, Q117M, Q117N, Q117P, Q117R, Q117S,Q117T, Q117V, Q117W, Q117Y, G118*, G118A, G118D, G118E, G118F, G118H,G118I, G118K, G118L, G118M, G118N, G118P, G118Q, G118R, G118S, G118T,G118V, G118W, G118Y, G119*, G119A, G119D, G119E, G119F, G119H, G119I,G119K, G119L, G119M, G119N, G119P, G119Q, G119R, G119S, G119T, G119V,G119W, G119Y, T120*, T120A, T120D, T120E, T120F, T120G, T120H, T120I,T120K, T120L, T120M, T120N, T120P, T120Q, T120R, T120S, T120V, T120W,T120Y, N121*, N121A, N121D, N121E, N121F, N121G, N121H, N121I, N121K,N121L, N121M, N121P, N121Q, N121R, N121S, N121T, N121V, N121W, N121Y,A122*, A122D, A122E, A122F, A122G, A122H, A122I, A122K, A122L, A122M,A122N, A122P, A122Q, A122R, A122S, A122T, A122V, A122W, A122Y, I123*,I123A, I123D, I123E, I123F, I123G, I123H, I123K, I123L, I123M, I123N,I123P, I123Q, I123R, I123S, I123T, I123V, I123W, I123Y, L124*, L124A,L124D, L124E, L124F, L124G, L124H, L124I, L124K, L124M, L124N, L124P,L124Q, L124R, L124S, L124T, L124V, L124W, L124Y, A125*, A125D, A125E,A125F, A125G, A125H, A125I, A125K, A125L, A125M, A125N, A125P, A125Q,A125R, A125S, A125T, A125V, A125W, A125Y, P126*, P126A, P126D, P126E,P126F, P126G, P126H, P126I, P126K, P126L, P126M, P126N, P126Q, P126R,P126S, P126T, P126V, P126W, P126Y, V127*, V127A, V127D, V127E, V127F,V127G, V127H, V127I, V127K, V127L, V127M, V127N, V127P, V127Q, V127R,V127S, V127T, V127W, V127Y, N128*, N128A, N128D, N128E, N128F, N128G,N128H, N128I, N128K, N128L, N128M, N128P, N128Q, N128R, N128S, N128T,N128V, N128W, N128Y, L129*, L129A, L129D, L129E, L129F, L129G, L129H,L129I, L129K, L129M, L129N, L129P, L129Q, L129R, L129S, L129T, L129V,L129W, L129Y, A130*, A130D, A130E, A130F, A130G, A130H, A130I, A130K,A130L, A130M, A130N, A130P, A130Q, A130R, A130S, A130T, A130V, A130W,A130Y, S131*, S131A, S131D, S131E, S131F, S131G, S131H, S131I, S131K,S131L, S131M, S131N, S131P, S131Q, S131R, S131T, S131V, S131W, S131Y,Q132*, Q132A, Q132D, Q132E, Q132F, Q132G, Q132H, Q132I, Q132K, Q132L,Q132M, Q132N, Q132P, Q132R, Q132S, Q132T, Q132V, Q132W, Q132Y, N133*,N133A, N133D, N133E, N133F, N133G, N133H, N133I, N133K, N133L, N133M,N133P, N133Q, N133R, N133S, N133T, N133V, N133W, N133Y, S134*, S134A,S134D, S134E, S134F, S134G, S134H, S134I, S134K, S134L, S134M, S134N,S134P, S134Q, S134R, S134T, S134V, S134W, S134Y, Q135*, Q135A, Q135D,Q135E, Q135F, Q135G, Q135H, Q135I, Q135K, Q135L, Q135M, Q135N, Q135P,Q135R, Q135S, Q135T, Q135V, Q135W, Q135Y, G136*, G136A, G136D, G136E,G136F, G136H, G136I, G136K, G136L, G136M, G136N, G136P, G136Q, G136R,G136S, G136T, G136V, G136W, G136Y, G137*, G137A, G137D, G137E, G137F,G137H, G137I, G137K, G137L, G137M, G137N, G137P, G137Q, G137R, G137S,G137T, G137V, G137W, G137Y, V138*, V138A, V138D, V138E, V138F, V138G,V138H, V138I, V138K, V138L, V138M, V138N, V138P, V138Q, V138R, V138S,V138T, V138W, V138Y, L139*, L139A, L139D, L139E, L139F, L139G, L139H,L139I, L139K, L139M, L139N, L139P, L139Q, L139R, L139S, L139T, L139V,L139W, L139Y, N140*, N140A, N140D, N140E, N140F, N140G, N140H, N140I,N140K, N140L, N140M, N140P, N140Q, N140R, N140S, N140T, N140V, N140W,N140Y, G141*, G141A, G141D, G141E, G141F, G141H, G141I, G141K, G141L,G141M, G141N, G141P, G141Q, G141R, G141S, G141T, G141V, G141W, G141Y,F142*, F142A, F142D, F142E, F142G, F142H, F142I, F142K, F142L, F142M,F142N, F142P, F142Q, F142R, F142S, F142T, F142V, F142W, F142Y, Y143*,Y143A, Y143D, Y143E, Y143F, Y143G, Y143H, Y143I, Y143K, Y143L, Y143M,Y143N, Y143P, Y143Q, Y143R, Y143S, Y143T, Y143V, Y143W, S144*, S144A,S144D, S144E, S144F, S144G, S144H, S144I, S144K, S144L, S144M, S144N,S144P, S144Q, S144R, S144T, S144V, S144W, S144Y, A145*, A145D, A145E,A145F, A145G, A145H, A145I, A145K, A145L, A145M, A145N, A145P, A145Q,A145R, A145S, A145T, A145V, A145W, A145Y, N146*, N146A, N146D, N146E,N146F, N146G, N146H, N146I, N146K, N146L, N146M, N146P, N146Q, N146R,N146S, N146T, N146V, N146W, N146Y, K147*, K147A, K147D, K147E, K147F,K147G, K147H, K147I, K147L, K147M, K147N, K147P, K147Q, K147R, K147S,K147T, K147V, K147W, K147Y, V148*, V148A, V148D, V148E, V148F, V148G,V148H, V148I, V148K, V148L, V148M, V148N, V148P, V148Q, V148R, V148S,V148T, V148W, V148Y, A149*, A149D, A149E, A149F, A149G, A149H, A149I,A149K, A149L, A149M, A149N, A149P, A149Q, A149R, A149S, A149T, A149V,A149W, A149Y, Q150*, Q150A, Q150D, Q150E, Q150F, Q150G, Q150H, Q150I,Q150K, Q150L, Q150M, Q150N, Q150P, Q150R, Q150S, Q150T, Q150V, Q150W,Q150Y, F151*, F151A, F151D, F151E, F151G, F151H, F151I, F151K, F151L,F151M, F151N, F151P, F151Q, F151R, F151S, F151T, F151V, F151W, F151Y,D152*, D152A, D152E, D152F, D152G, D152H, D152I, D152K, D152L, D152M,D152N, D152P, D152Q, D152R, D152S, D152T, D152V, D152W, D152Y, P153*,P153A, P153D, P153E, P153F, P153G, P153H, P153I, P153K, P153L, P153M,P153N, P153Q, P153R, P153S, P153T, P153V, P153W, P153Y, S154*, S154A,S154D, S154E, S154F, S154G, S154H, S154I, S154K, S154L, S154M, S154N,S154P, S154Q, S154R, S154T, S154V, S154W, S154Y, K155*, K155A, K155D,K155E, K155F, K155G, K155H, K155I, K155L, K155M, K155N, K155P, K155Q,K155R, K155S, K155T, K155V, K155W, K155Y, P156*, P156A, P156D, P156E,P156F, P156G, P156H, P156I, P156K, P156L, P156M, P156N, P156Q, P156R,P156S, P156T, P156V, P156W, P156Y, Q157*, Q157A, Q157D, Q157E, Q157F,Q157G, Q157H, Q157I, Q157K, Q157L, Q157M, Q157N, Q157P, Q157R, Q157S,Q157T, Q157V, Q157W, Q157Y, Q158*, Q158A, Q158D, Q158E, Q158F, Q158G,Q158H, Q158I, Q158K, Q158L, Q158M, Q158N, Q158P, Q158R, Q158S, Q158T,Q158V, Q158W, Q158Y, T159*, T159A, T159D, T159E, T159F, T159G, T159H,T159I, T159K, T159L, T159M, T159N, T159P, T159Q, T159R, T159S, T159V,T159W, T159Y, K160*, K160A, K160D, K16E, K160F, K160G, K160H, K160I,K160L, K165M, K160N, K165P, K160Q, K160R, K160S, K160T, K160V, K160W,K160Y, G161*, G161A, G161D, G161E, G161F, G161H, G161I, G161K, G161L,G161M, G161N, G161P, G161Q, G161R, G161S, G161T, G161V, G161W, G161Y,T162A, T162D, T162E, T162F, T162G, T162H, T162I, T162K, T162L, T162M,T162N, T162P, T162Q, T162R, T162S, T162T, T162V, T162W, T162Y, W163*,W163A, W163D, W163E, W163F, W163G, W163H, W163I, W163K, W163L, W163M,W163N, W163P, W163Q, W163R, W163S, W163T, W163V, W163Y, F164*, F164A,F164D, F164E, F164G, F164H, F164I, F164K, F164L, F164M, F164N, F164P,F164Q, F164R, F164S, F164T, F164V, F164W, F164Y, Q165*, Q165A, Q165D,Q165E, Q165F, Q165G, Q165H, Q165I, Q165K, Q165L, Q165M, Q165N, Q165P,Q165R, Q165S, Q165T, Q165V, Q165W, Q165Y, I166*, I166A, I166D, I166E,I166F, I166G, I166H, I166K, I166L, I166M, I166N, I166P, I166Q, I166R,I166S, I166T, I166V, I166W, I166Y, T167*, T167A, T167D, T167E, T167F,T167G, T167H, T167I, T167K, T167L, T167M, T167N, T167P, T167Q, T167R,T167S, T167V, T167W, T167Y, K168*, K168A, K168D, K168E, K168F, K168G,K168H, K168I, K168L, K168M, K168N, K168P, K168Q, K168R, K168S, K168T,K168V, K168W, K168Y, F169*, F169A, F169D, F169E, F169G, F169H, F169I,F169K, F169L, F169M, F169N, F169P, F169Q, F169R, F169S, F169T, F169V,F169W, F169Y, T170*, T170A, T170D, T170E, T170F, T170G, T170H, T170I,T170K, T170L, T170M, T170N, T170P, T170Q, T170R, T170S, T170V, T170W,T170Y, G171*, G171A, G171D, G171E, G171F, G171H, G171I, G171K, G171L,G171M, G171N, G171P, G171Q, G171R, G171S, G171T, G171V, G171W, G171Y,A172*, A172D, A172E, A172F, A172G, A172H, A172I, A172K, A172L, A172M,A172N, A172P, A172Q, A172R, A172S, A172T, A172V, A172W, A172Y, A173*,A173D, A173E, A173F, A173G, A173H, A173I, A173K, A173L, A173M, A173N,A173P, A173Q, A173R, A173S, A173T, A173V, A173W, A173Y, G174*, G174A,G174D, G174E, G174F, G174H, G174I, G174K, G174L, G174M, G174N, G174P,G174Q, G174R, G174S, G174T, G174V, G174W, G174Y, P175*, P175A, P175D,P175E, P175F, P175G, P175H, P175I, P175K, P175L, P175M, P175N, P175Q,P175R, P175S, P175T, P175V, P175W, P175Y, Y176*, Y176A, Y176D, Y176E,Y176F, Y176G, Y176H, Y176I, Y176K, Y176L, Y176M, Y176N, Y176P, Y176Q,Y176R, Y176S, Y176T, Y176V, Y176W, K178*, K178A, K178D, K178E, K178F,K178G, K178H, K178I, K178L, K178M, K178N, K178P, K178Q, K178R, K178S,K178T, K178V, K178W, K178Y, A179*, A179D, A179E, A179F, A179G, A179H,A179I, A179K, A179L, A179M, A179N, A179P, A179Q, A179R, A179S, A179T,A179V, A179W, A179Y, L180*, L180A, L180D, L180E, L180F, L180G, L180H,L180I, L180K, L180M, L180N, L180P, L180Q, L180R, L180S, L180T, L180V,L180W, L180Y, G181*, G181A, G181D, G181E, G181F, G181H, G181I, G181K,G181L, G181M, G181N, G181P, G181Q, G181R, G181S, G181T, G181V, G181W,G181Y, S182*, S182A, S182D, S182E, S182F, S182G, S182H, S182I, S182K,S182L, S182M, S182N, S182P, S182Q, S182R, S182T, S182V, S182W, S182Y,N183*, N183A, N183D, N183E, N183F, N183G, N183H, N183I, N183K, N183L,N183M, N183P, N183Q, N183R, N183S, N183T, N183V, N183W, N183Y, D184*,D184A, D184E, D184F, D184G, D184H, D184I, D184K, D184L, D184M, D184N,D184P, D184Q, D184R, D184S, D184T, D184V, D184W, D184Y, K185*, K185A,K185D, K185E, K185F, K185G, K185H, K185I, K185L, K185M, K185N, K185P,K185Q, K185R, K185S, K185T, K185V, K185W, K185Y, S186*, S186A, S186D,S186E, S186F, S186G, S186H, S186I, S186K, S186L, S186M, S186N, S186P,S186Q, S186R, S186T, S186V, S186W, S186Y, V187*, V187A, V187D, V187E,V187F, V187G, V187H, V187I, V187K, V187L, V187M, V187N, V187P, V187Q,V187R, V187S, V187T, V187W, V187Y, D189*, D189A, D189E, D189F, D189G,D189H, D189I, D189K, D189L, D189M, D189N, D189P, D189Q, D189R, D189S,D189T, D189V, D189W, D189Y, K190*, K190A, K190D, K190E, K190F, K190G,K190H, K190I, K190L, K190M, K190N, K190P, K190Q, K190R, K190S, K190T,K190V, K190W, K190Y, N191*, N191A, N191D, N191E, N191F, N191G, N191H,N191I, N191K, N191L, N191M, N191P, N191Q, N191R, N191S, N191T, N191V,N191W, N191Y, K192*, K192A, K192D, K192E, K192F, K192G, K192H, K192I,K192L, K192M, K192N, K192P, K192Q, K192R, K192S, K192T, K192V, K192W,K192Y, N193*, N193A, N193D, N193E, N193F, N193G, N193H, N193I, N193K,N193L, N193M, N193P, N193Q, N193R, N193S, N193T, N193V, N193W, N193Y,I194*, I194A, I194D, I194E, I194F, I194G, I194H, I194K, I194L, I194M,I194N, I194P, I194Q, I194R, I194S, I194T, I194V, I194W, I194Y, A195*,A195D, A195E, A195F, A195G, A195H, A195I, A195K, A195L, A195M, A195N,A195P, A195Q, A195R, A195S, A195T, A195V, A195W, A195Y, G196*, G196A,G196D, G196E, G196F, G196H, G196I, G196K, G196L, G196M, G196N, G196P,G196Q, G196R, G196S, G196T, G196V, G196W, G196Y, D197*, D197A, D197E,D197F, D197G, D197H, D197I, D197K, D197L, D197M, D197N, D197P, D197Q,D197R, D197S, D197T, D197V, D197W, D197Y, W198*, W198A, W198D, W198E,W198F, W198G, W198H, W198I, W198K, W198L, W198M, W198N, W198P, W198Q,W198R, W198S, W198T, W198V, W198Y, G199*, G199A, G199D, G199E, G199F,G199H, G199I, G199K, G199L, G199M, G199N, G199P, G199Q, G199R, G199S,G199T, G199V, G199W, G199Y, F200*, F200A, F200D, F200E, F200G, F200H,F200I, F200K, F200L, F200M, F200N, F200P, F200Q, F200R, F200S, F200T,F200V, F200W, F200Y, D201*, D201A, D201E, D201F, D201G, D201H, D201I,D201K, D201L, D201M, D201N, D201P, D201Q, D201R, D201S, D201T, D201V,D201W, D201Y, P202*, P202A, P202D, P202E, P202F, P202G, P202H, P202I,P202K, P202L, P202M, P202N, P202Q, P202R, P202S, P202T, P202V, P202W,P202Y, A203*, A203D, A203E, A203F, A203G, A203H, A203I, A203K, A203L,A203M, A203N, A203P, A203Q, A203R, A203S, A203T, A203V, A203W, A203Y,K204*, K204A, K204D, K204E, K204F, K204G, K204H, K204I, K204L, K204M,K204N, K204P, K204Q, K204R, K204S, K204T, K204V, K204W, K204Y, W205*,W205A, W205D, W205F, W205F, W205G, W205H, W205I, W205K, W205I, W205M,W205N, W205P, W205Q, W205R, W205S, W205T, W205V, W205Y, A206*, A206D,A206E, A206F, A206G, A206H, A206I, A206K, A206L, A206M, A206N, A206P,A206Q, A206R, A206S, A206T, A206V, A206W, A206Y, Y207*, Y207A, Y207D,Y207E, Y207F, Y207G, Y207H, Y207I, Y207K, Y207L, Y207M, Y207N, Y207P,Y207Q, Y207R, Y207S, Y207T, Y207V, Y207W, Q208*, Q208A, Q208D, Q208E,Q208F, Q208G, Q208H, Q208I, Q208K, Q208L, Q208M, Q208N, Q208P, Q208R,Q208S, Q208T, Q208V, Q208W, Q208Y, Y209*, Y209A, Y209D, Y209E, Y209F,Y209G, Y209H, Y209I, Y209K, Y209L, Y209M, Y209N, Y209P, Y209Q, Y209R,Y209S, Y209T, Y209V, Y209W, D210*, D210A, D210E, D210F, D210G, D210H,D210I, D210K, D210L, D210M, D210N, D210P, D210Q, D210R, D210S, D210T,D210V, D210W, D210Y, E211*, E211A, E211D, E211F, E211G, E211H, E211I,E211K, E211L, E211M, E211N, E211P, E211Q, E211R, E211S, E211T, E211V,E211W, E211Y, K212*, K212A, K212D, K212E, K212F, K212G, K212H, K212I,K212L, K212M, K212N, K212P, K212Q, K212R, K212S, K212T, K212V, K212W,K212Y, N213*, N213A, N213D, N213E, N213F, N213G, N213H, N213I, N213K,N213L, N213M, N213P, N213Q, N213R, N213S, N213T, N213V, N213W, N213Y,N214*, N214A, N214D, N214E, N214F, N214G, N214H, N214I, N214K, N214L,N214M, N214P, N214Q, N214R, N214S, N214T, N214V, N214W, N214Y, K215*,K215A, K215D, K215E, K215F, K215G, K215H, K215I, K215L, K215M, K215N,K215P, K215Q, K215R, K215S, K215T, K215V, K215W, K215Y, F216*, F216A,F216D, F216E, F216G, F216H, F216I, F216K, F216L, F216M, F216N, F216P,F216Q, F216R, F216S, F216T, F216V, F216W, F216Y, N217*, N217A, N217D,N217E, N217F, N217G, N217H, N217I, N217K, N217L, N217M, N217P, N217Q,N217R, N217S, N217T, N217V, N217W, N217Y, Y218*, Y218A, Y218D, Y218E,Y218F, Y218G, Y218H, Y218I, Y218K, Y218L, Y218M, Y218N, Y218P, Y218Q,Y218R, Y218S, Y218T, Y218V, Y218W, V219*, V219A, V219D, V219E, V219F,V219G, V219H, V219I, V219K, V219L, V219M, V219N, V219P, V219Q, V219R,V219S, V219T, V219W, V219Y, G220*, G220A, G220D, G220E, G220F, G220H,G220I, G220K, G220L, G220M, G220N, G220P, G220Q, G220R, G220S, G220T,G220V, G220W, G220Y, K221*, K221A, K221D, K221E, K221F, K221G, K221H,K221I, K221L, K221M, K221N, K221P, K221Q, K221R, K221S, K221T, K221V,K221W or K221Y, wherein the variant has an amino acid sequence which isat least 60%, at least 70%, at least 75%, at least 80%, at least 85%, atleast 90% or at least 95% identical to SEQ ID NO: 1.

The present invention also relates to isolated polynucleotides encodingthe variants; nucleic acid constructs, vectors, and host cellscomprising the polynucleotides; and methods of producing the variants.The invention further relates to detergent compositions comprising suchvariants.

Definitions

DNase (deoxyribonuclease): The term “DNase” means a polypeptide withDNase activity that catalyzes the hydrolytic cleavage of phosphodiesterlinkages in DNA, thus degrading DNA. DNases belong to the esterases(EC-number 3.1), a subgroup of the hydrolases. The DNases are classifiedEC 3.1.21.1. For purposes of the present invention, DNase activity isdetermined according to the procedure described in the Assay I. In oneaspect, the DNase variants of the present invention have improved DNaseactivity compared to the parent DNase. In one aspect, the DNase variantsof the present invention have at least 100%, e.g., at least 110%, atleast 120%, at least 130%, at least 140%, at least 150%, at least 160%,at least 170%, at least 180%, at least 190%, or at least 200% DNaseactivity compared to the polypeptide with SEQ ID NO: 1.

The term “parent”: the term parent means any polypeptide with DNaseactivity to which an alteration is made to produce the DNase variants ofthe invention. Thus a DNase parent or precursor DNase means a DNase inwhich an alteration is made to produce the DNase variants of the presentinvention. The terms parent and precursor may be used interchangeably inthe present application. Thus, the parent is a DNase having theidentical amino acid sequence of the variant but not having thealterations at one or more of the specified positions. It will beunderstood, that in the present context the expression “having identicalamino acid sequence” relates to 100% sequence identity. In a particularembodiment the DNase parent is a DNase with at least 60%, at least 61%,at least 62%, at least 63%, at least 64%, at least 65%, at least 66%, atleast 67%, at least 68%, at least 69%, at least 70%, at least 72%, atleast 73%, at least 74%, at least 75%, at least 80%, at least 81%, atleast 82%, at least 83%, at least 84%, at least 85%, at least 90%, atleast 91%, at least 92%, at least 93%, at least 94%, at least 95%, atleast 96%, at least 97%, at least 98%, at least 99%, e. g. at least99.1%, at least 99.2%, at least 99.3%, at least 99.4%, at least 99.5%,at least 99.6 or 100% identity to a polypeptide with SEQ ID NO: 1.

The term “DNase variant” means a DNase having DNase activity and whichcomprises an alteration, i.e., a substitution, insertion, and/ordeletion at one or more (or one or several) positions compared to itsparent e.g. compared to SEQ ID NO: 1. A substitution means a replacementof an amino acid occupying a position with a different amino acid; adeletion means removal of an amino acid occupying a position; and aninsertion means adding amino acids e.g. 1 to 10 amino acids, preferably1-3 amino acids adjacent to an amino acid occupying a position.Preferably, the variant is modified by the hand of man. In one aspect,the variant is at least 1% pure, e.g., at least 5% pure, at least 10%pure, at least 20% pure, at least 40% pure, at least 60% pure, at least80% pure, and at least 90% pure, as determined by SDS PAGE.

The term “allelic variant” means any of two or more alternative forms ofa gene occupying the same chromosomal locus. Allelic variation arisesnaturally through mutation, and may result in polymorphism withinpopulations. Gene mutations can be silent (no change in the encodedpolypeptide) or may encode polypeptides having altered amino acidsequences. An allelic variant of a polypeptide is a polypeptide encodedby an allelic variant of a gene.

The term “isolated polynucleotide” means a polynucleotide that ismodified by the hand of man. In one aspect, the isolated polynucleotideis at least 1% pure, e.g., at least 5% pure, at least 10% pure, at least20% pure, at least 40% pure, at least 60% pure, at least 80% pure, atleast 90% pure, and at least 95% pure, as determined by agaroseelectrophoresis. The polynucleotides may be of genomic, cDNA, RNA,semisynthetic, synthetic origin, or any combinations thereof.

The term “substantially pure variant” means a preparation that containsat most 10%, at most 8%, at most 6%, at most 5%, at most 4%, at most 3%,at most 2%, at most 1%, and at most 0.5% by weight of other polypeptidematerial with which it is natively or recombinantly associated.Preferably, the variant is at least 92% pure, e.g., at least 94% pure,at least 95% pure, at least 96% pure, at least 97% pure, at least 98%pure, at least 99%, at least 99.5% pure, and 100% pure by weight of thetotal polypeptide material present in the preparation. The variants ofthe present invention are preferably in a substantially pure form. Thiscan be accomplished, for example, by preparing the variant by well-knownrecombinant methods or by classical purification methods.

The term “wild-type DNase” means a DNase expressed by a naturallyoccurring organism, such as a fungal, bacterium, archaea, yeast, plantor animal found in nature.

The term “mature polypeptide” means a polypeptide in its final formfollowing translation and any post-translational modifications, such asN-terminal processing, C-terminal truncation, glycosylation,phosphorylation, etc. In one aspect, the mature polypeptide correspondsto the amino acid sequence with SEQ ID NO: 1.

The term “mature polypeptide coding sequence” means a polynucleotidethat encodes a mature polypeptide having DNase activity.

The term “cDNA” means a DNA molecule that can be prepared by reversetranscription from a mature, spliced, mRNA molecule obtained from aprokaryotic or eukaryotic cell. A cDNA lacks intron sequences that maybe present in the corresponding genomic DNA. The initial, primary RNAtranscript is a precursor to mRNA that is processed through a series ofsteps, including splicing, before appearing as mature spliced mRNA.

The term “coding sequence” means a polynucleotide, which directlyspecifies the amino acid sequence of its polypeptide product. Theboundaries of the coding sequence are generally determined by an openreading frame, which usually begins with the ATG start codon oralternative start codons such as GTG and TTG and ends with a stop codonsuch as TAA, TAG, and TGA. The coding sequence may be a DNA, cDNA,synthetic, or recombinant polynucleotide.

The term “nucleic acid construct” means a nucleic acid molecule, eithersingle- or double-stranded, which is isolated from a naturally occurringgene or is modified to contain segments of nucleic acids in a mannerthat would not otherwise exist in nature or which is synthetic. The termnucleic acid construct is synonymous with the term “expression cassette”when the nucleic acid construct contains the control sequences requiredfor expression of a coding sequence of the present invention.

The term “operably linked” means a configuration in which a controlsequence is placed at an appropriate position relative to the codingsequence of a polynucleotide such that the control sequence directs theexpression of the coding sequence.

The term “control sequences” means all components necessary for theexpression of a polynucleotide encoding a variant of the presentinvention. Each control sequence may be native or foreign to thepolynucleotide encoding the variant or native or foreign to each other.Such control sequences include, but are not limited to, a leader,polyadenylation sequence, propeptide sequence, promoter, signal peptidesequence, and transcription terminator. At a minimum, the controlsequences include a promoter, and transcriptional and translational stopsignals. The control sequences may be provided with linkers for thepurpose of introducing specific restriction sites facilitating ligationof the control sequences with the coding region of the polynucleotideencoding a variant.

The term “expression” includes any step involved in the production ofthe variant including, but not limited to, transcription,post-transcriptional modification, translation, post-translationalmodification, and secretion.

The term “expression vector” means a linear or circular DNA moleculethat comprises a polynucleotide encoding a variant and is operablylinked to additional nucleotides that provide for its expression.

The term “transcription promoter” is used for a promoter which is aregion of DNA that facilitates the transcription of a particular gene.Transcription promoters are typically located near the genes theyregulate, on the same strand and upstream (towards the 5′ region of thesense strand).

The term “transcription terminator” is used for a section of the geneticsequence that marks the end of gene or operon on genomic DNA fortranscription.

The term “host cell” means any cell type that is susceptible totransformation, transfection, transduction, and the like with a nucleicacid construct or expression vector comprising a polynucleotide of thepresent invention. The term “host cell” encompasses any progeny of aparent cell that is not identical to the parent cell due to mutationsthat occur during replication.

The term “high stringency conditions” means for probes of at least 100nucleotides in length, prehybridization and hybridization at 42° C. in5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon spermDNA, and 50% formamide, following standard Southern blotting proceduresfor 12 to 24 hours. The carrier material is finally washed three timeseach for 15 minutes using 2×SSC, 0.2% SDS at 65° C.

The term “very high stringency conditions” means for probes of at least100 nucleotides in length, prehybridization and hybridization at 42° C.in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmonsperm DNA, and 50% formamide, following standard Southern blottingprocedures for 12 to 24 hours. The carrier material is finally washedthree times each for 15 minutes using 2×SSC, 0.2% SDS at 70° C.

The term “medium stringency conditions” means for probes of at least 100nucleotides in length, prehybridization and hybridization at 42° C. in5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon spermDNA, and 35% formamide, following standard Southern blotting proceduresfor 12 to 24 hours. The carrier material is finally washed three timeseach for 15 minutes using 2×SSC, 0.2% SDS at 55° C.

The term “medium-high stringency conditions” means for probes of atleast 100 nucleotides in length, prehybridization and hybridization at42° C. in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denaturedsalmon sperm DNA, and either 35% formamide, following standard Southernblotting procedures for 12 to 24 hours. The carrier material is finallywashed three times each for 15 minutes using 2×SSC, 0.2% SDS at 60° C.

The term “low stringency conditions” means for probes of at least 100nucleotides in length, prehybridization and hybridization at 42° C. in5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmon spermDNA, and 25% formamide, following standard Southern blotting proceduresfor 12 to 24 hours. The carrier material is finally washed three timeseach for 15 minutes using 2×SSC, 0.2% SDS at 50° C.

The term “very low stringency conditions” means for probes of at least100 nucleotides in length, prehybridization and hybridization at 42° C.in 5×SSPE, 0.3% SDS, 200 micrograms/ml sheared and denatured salmonsperm DNA, and 25% formamide, following standard Southern blottingprocedures for 12 to 24 hours. The carrier material is finally washedthree times each for 15 minutes using 2×SSC, 0.2% SDS at 45° C.

The term “improved property” means a characteristic associated with avariant that is improved compared to the parent and/or compared to aDNase with SEQ ID NO: 1, or compared to a DNase having the identicalamino acid sequence of said variant but not having the alterations atone or more of said specified positions. Such improved propertiesinclude, but are not limited to, stability, such as detergent stability,wash performance e.g. deep cleaning effect, the term “deep cleaning” ismeant disruption or removal of a biofilm or components of a biofilm suchas polysaccharides, proteins, DNA, soil or other components present inthe biofilm.

A biofilm is any group of microorganisms in which cells stick to eachother on a surface, such as a textile, dishware or hard surface. Theseadherent cells are frequently embedded within a self-produced matrix ofextracellular polymeric substance (EPS). Biofilm EPS is a polymericconglomeration generally composed of extracellular DNA, proteins, andpolysaccharides. Biofilms may form on living or non-living surfaces. Themicrobial cells growing in a biofilm are physiologically distinct fromplanktonic cells of the same organism, which, by contrast, aresingle-cells that may float or swim in a liquid medium. Bacteria livingin a biofilm usually have significantly different properties fromfree-floating bacteria of the same species, as the dense and protectedenvironment of the film allows them to cooperate and interact in variousways. One benefit of this environment is increased resistance todetergents and antibiotics, as the dense extracellular matrix and theouter layer of cells protect the interior of the community. On laundrybiofilm producing bacteria can be found among the following species:Acinetobacter sp., Aeromicrobium sp., Brevundimonas sp., Microbacteriumsp., Micrococcus luteus, Pseudomonas sp., Staphylococcus epidermidis,and Stenotrophomonas sp.

The term “improved DNase activity” is defined herein as an altered DNaseactivity e.g. by increased catalyse of hydrolytic cleavage ofphosphodiester linkages in the DNA the DNase variant displaying analteration of the activity relative (or compared) to the activity of theparent DNase, such as compared to a DNase with SEQ ID NO: 1.

The term “stability” includes storage stability and stability duringuse, e.g. during a wash process and reflects the stability of the DNasevariant according to the invention as a function of time e.g. how muchactivity is retained when the DNase variant is kept in solution inparticular in a detergent solution. The stability is influenced by manyfactors e.g. pH, temperature, detergent composition e.g. amount ofbuilder, surfactants etc. The DNase stability may be measured asdescribed in example 2. The term “improved stability” or “increasedstability” is defined herein as a variant DNase displaying an increasedstability in solutions, relative to the stability of the parent DNaseand/or relative to SEQ ID NO: 1. “Improved stability” and “increasedstability” includes detergent stability. The term “detergent stability”or “improved detergent stability may be improved stability of the DNaseactivity compared to the DNase parent. The DNase stability is measuredas described in example 2.

The term “improved wash performance” may be defined as improved deepcleaning effect i.e. the disruption or removal of a biofilm orcomponents of a DNase variant according to the invention compared to theDNase parent or the DNase with SEQ ID NO: 1. The DNase variants may alsohave improved malodor removal. By the term“malodor” is meant an odorwhich is not desired on clean items. The cleaned item should smell freshand clean without malodors adhered to the item. One example of malodoris compounds with an unpleasant smell, which may be produced bymicroorganisms. Another example is unpleasant smells can be sweat orbody odor adhered to an item which has been in contact with human oranimal. Another example of malodor can be the odor from spices, whichsticks to items for example curry or other exotic spices which smellsstrongly. One way of measuring the ability of an item to adhere malodoris by using Assay II disclosed herein.

Wash performance may be expressed as a Remission value of the stainedswatches. After washing and rinsing the swatches are spread out flat andallowed to air dry at room temperature overnight. All washed swatchesare evaluated the day after the wash. Light reflectance evaluations ofthe swatches are done using a Macbeth Color Eye 7000 reflectancespectrophotometer with very small aperture. The measurements are madewithout UV in the incident light and remission at 460 nm was extracted.

The term “laundering” relates to both household laundering andindustrial laundering and means the process of treating textiles with asolution containing a cleaning or detergent composition of the presentinvention. The laundering process can for example be carried out usinge.g. a household or an industrial washing machine or can be carried outby hand.

The term “detergent composition”, includes unless otherwise indicated,granular or powder-form all-purpose or heavy-duty washing agents,especially cleaning detergents; liquid, gel or paste-form all-purposewashing agents, especially the so-called heavy-duty liquid (HDL) types;liquid fine-fabric detergents; hand dishwashing agents or light dutydishwashing agents, especially those of the high-foaming type; machinedishwashing agents, including the various tablet, granular, liquid andrinse-aid types for household and institutional use; liquid cleaning anddisinfecting agents, including antibacterial hand-wash types, cleaningbars, soap bars, mouthwashes, denture cleaners, car or carpet shampoos,bathroom cleaners; hair shampoos and hair-rinses; shower gels, foambaths; metal cleaners; as well as cleaning auxiliaries such as bleachadditives and “stain-stick” or pre-treat types. The terms “detergentcomposition” and “detergent formulation” are used in reference tomixtures which are intended for use in a wash medium for the cleaning ofsoiled objects. In some embodiments, the term is used in reference tolaundering fabrics and/or garments (e.g., “laundry detergents”). Inalternative embodiments, the term refers to other detergents, such asthose used to clean dishes, cutlery, etc. (e.g., “dishwashingdetergents”). It is not intended that the present invention be limitedto any particular detergent formulation or composition. The term“detergent composition” is not intended to be limited to compositionsthat contain surfactants. It is intended that in addition to thevariants according to the invention, the term encompasses detergentsthat may contain, e.g., surfactants, builders, chelators or chelatingagents, bleach system or bleach components, polymers, fabricconditioners, foam boosters, suds suppressors, dyes, perfume, tannishinhibitors, optical brighteners, bactericides, fungicides, soilsuspending agents, anti-corrosion agents, enzyme inhibitors orstabilizers, enzyme activators, transferase(s), hydrolytic enzymes,oxido reductases, bluing agents and fluorescent dyes, antioxidants, andsolubilizers.

The term “fabric” encompasses any textile material. Thus, it is intendedthat the term encompass garments, as well as fabrics, yarns, fibers,non-woven materials, natural materials, synthetic materials, and anyother textile material.

The term “textile” refers to woven fabrics, as well as staple fibers andfilaments suitable for conversion to or use as yarns, woven, knit, andnon-woven fabrics. The term encompasses yarns made from natural, as wellas synthetic (e.g., manufactured) fibers. The term, “textile materials”is a general term for fibers, yarn intermediates, yarn, fabrics, andproducts made from fabrics (e.g., garments and other articles).

The term “non-fabric detergent compositions” include non-textile surfacedetergent compositions, including but not limited to compositions forhard surface cleaning, such as dishwashing detergent compositionsincluding manual dish wash compositions, oral detergent compositions,denture detergent compositions, and personal cleansing compositions.

The term “effective amount of enzyme” refers to the quantity of enzymenecessary to achieve the enzymatic activity required in the specificapplication, e.g., in a defined detergent composition. Such effectiveamounts are readily ascertained by one of ordinary skill in the art andare based on many factors, such as the particular enzyme used, thecleaning application, the specific composition of the detergentcomposition, and whether a liquid or dry (e.g., granular, bar)composition is required, and the like. The term “effective amount” of aDNase variant refers to the quantity of DNase variant describedhereinbefore that achieves a desired level of enzymatic activity, e.g.,in a defined detergent composition.

The term “water hardness” or “degree of hardness” or “dH” or “° dH” asused herein refers to German degrees of hardness. One degree is definedas 10 milligrams of calcium oxide per liter of water.

The term “relevant washing conditions” is used herein to indicate theconditions, particularly washing temperature, time, washing mechanics,detergent concentration, type of detergent and water hardness, actuallyused in households in a detergent market segment.

The term “adjunct materials” means any liquid, solid or gaseous materialselected for the particular type of detergent composition desired andthe form of the product (e.g., liquid, granule, powder, bar, paste,spray, tablet, gel, or foam composition), which materials are alsopreferably compatible with the DNase variant enzyme used in thecomposition. In some embodiments, granular compositions are in “compact”form, while in other embodiments, the liquid compositions are in a“concentrated” form.

The term “low detergent concentration” system includes detergents whereless than about 800 ppm of detergent components is present in the washwater. Asian, e.g., Japanese detergents are typically considered lowdetergent concentration systems.

The term “medium detergent concentration” system includes detergentswherein between about 800 ppm and about 2000 ppm of detergent componentsis present in the wash water. North American detergents are generallyconsidered to be medium detergent concentration systems.

The term “high detergent concentration” system includes detergentswherein greater than about 2000 ppm of detergent components is presentin the wash water. European detergents are generally considered to behigh detergent concentration systems.

Conventions for Designation of Variants

For purposes of the present invention, the mature polypeptide disclosedin SEQ ID NO: 1 is used to determine the corresponding amino acidresidue in another DNase. The amino acid sequence of another DNase isaligned with the polypeptide disclosed in SEQ ID NO: 1, and based on thealignment, the amino acid position number corresponding to any aminoacid residue in the polypeptide disclosed in SEQ ID NO: 1 is determinedusing the Needleman-Wunsch algorithm (Needleman and Wunsch, 1970, J.Mol. Biol. 48: 443-453) as implemented in the Needle program of theEMBOSS package (EMBOSS: The European Molecular Biology Open SoftwareSuite, Rice et al., 2000, Trends Genet. 16: 276-277), preferably version5.0.0 or later. The parameters used are gap open penalty of 10, gapextension penalty of 0.5, and the EBLOSUM62 (EMBOSS version of BLOSUM62)substitution matrix.

Identification of the corresponding amino acid residue in another DNasecan be determined by an alignment of multiple polypeptide sequencesusing several computer programs including, but not limited to, MUSCLE(multiple sequence comparison by log-expectation; version 3.5 or later;Edgar, 2004, Nucleic Acids Research 32: 1792-1797), MAFFT (version 6.857or later; Katoh and Kuma, 2002, Nucleic Acids Research 30: 3059-3066;Katoh et al., 2005, Nucleic Acids Research 33: 511-518; Katoh and Toh,2007, Bioinformatics 23: 372-374; Katoh et al., 2009, Methods inMolecular Biology 537:_39-64; Katoh and Toh, 2010, Bioinformatics26:_1899-1900), and EMBOSS EMMA employing ClustalW (1.83 or later;Thompson et al., 1994, Nucleic Acids Research 22: 4673-4680), usingtheir respective default parameters.

When the other enzyme has diverged from the polypeptide of SEQ ID NO: 1such that traditional sequence-based comparison fails to detect theirrelationship (Lindahl and Elofsson, 2000, J. Mol. Biol. 295: 613-615),other pairwise sequence comparison algorithms can be used. Greatersensitivity in sequence-based searching can be attained using searchprograms that utilize probabilistic representations of polypeptidefamilies (profiles) to search databases. For example, the PSI-BLASTprogram generates profiles through an iterative database search processand is capable of detecting remote homologs (Atschul et al., 1997,Nucleic Acids Res. 25: 3389-3402). Even greater sensitivity can beachieved if the family or superfamily for the polypeptide has one ormore representatives in the protein structure databases. Programs suchas GenTHREADER (Jones, 1999, J. Mol. Biol. 287: 797-815; McGuffin andJones, 2003, Bioinformatics 19: 874-881) utilize information from avariety of sources (PSI-BLAST, secondary structure prediction,structural alignment profiles, and solvation potentials) as input to aneural network that predicts the structural fold for a query sequence.Similarly, the method of Gough et al., 2000, J. Mol. Biol. 313: 903-919,can be used to align a sequence of unknown structure with thesuperfamily models present in the SCOP database. These alignments can inturn be used to generate homology models for the polypeptide, and suchmodels can be assessed for accuracy using a variety of tools developedfor that purpose.

For proteins of known structure, several tools and resources areavailable for retrieving and generating structural alignments. Forexample, the SCOP super families of proteins have been structurallyaligned, and those alignments are accessible and downloadable. Two ormore protein structures can be aligned using a variety of algorithmssuch as the distance alignment matrix (Holm and Sander, 1998, Proteins33: 88-96) or combinatorial extension (Shindyalov and Bourne, 1998,Protein Engineering 11: 739-747), and implementation of these algorithmscan additionally be utilized to query structure databases with astructure of interest in order to discover possible structural homologs(e.g., Holm and Park, 2000, Bioinformatics 16: 566-567).

In describing the variants of the present invention, the nomenclaturedescribed below is adapted for ease of reference. The accepted IUPACsingle letter or three letters amino acid abbreviations are employed.Amino acid positions are indicated with #₁, #₂, etc.

Substitutions: For an amino acid substitution, the followingnomenclature is used: Original amino acid, position, substituted aminoacid. Accordingly, the substitution of valine at position #₁ withalanine is designated as “Val #₁Ala” or “V #₁A”. Multiple mutations areseparated by addition marks (“+”) or by commas (,), e.g., “Val#₁Ala+“Pro #₂Gly” or V #₁A, P #₂G, representing substitutions atpositions #₁ and #₂ of valine (V) and proline with alanine (A) andglycine (G), respectively. If more than one amino acid may besubstituted in a given position these are listed in brackets, such as[X] or {X} or alternatively separated by commas. Thus if both Trp andLys according to the invention may be substituted instead of the aminoacid occupying at position #₁ this is indicated as X #₁{W, K}, X #₁[W,K] or X #₁ W, K; where the X indicate the amino acid residues ofdifferent DNases which according to the invention may be the parentDNase e.g. such as a DNase with SEQ ID NO: 1 or a DNase having at least60% identity hereto. Thus in some cases the variants are represented as#₁{W, K} or X #₂P indicating that the amino acids to be substituted varydepending on the parent. For convenience as SEQ ID NO: 1 is used fornumbering the substitutions the amino acid in the corresponding positionin SEQ ID NO: 1 is indicated e.g. V1A. However, it will be clear to theskilled artisan that a DNase variant comprising V1A is not limited toparent DNases having valine at a position corresponding to position 1 ofSEQ ID NO: 1. In a parent DNase having e.g. asparagine in position 1,the skilled person would translate the mutation specification V1A toN1A. In the event the parent DNase has alanine in position 1, theskilled person would recognize that the parent DNase is not changed atthis position.

Deletions: For an amino acid deletion, the following nomenclature isused: Original amino acid, position, *. Accordingly, the deletion ofserine at position #₁ is designated as “Ser #₁*” or “S #₁*”. Multipledeletions are separated by addition marks (“+”) or commas, e.g., “Val#₁*+Pro #₂*” or “V #₁*, P #₂*”.

Insertions: The insertion of an additional amino acid residue such ase.g. a lysine after Val #, may be indicated by: Val #₁ValLys or V #₁VK.Alternatively insertion of an additional amino acid residue such aslysine after V #₁ may be indicated by: *#₁aK. When more than one aminoacid residue is inserted, such as e.g. a Lys, and Gly after #₁ this maybe indicated as: Ala #₁AlaLysGly or A #₁AKG. In such cases, the insertedamino acid residue(s) may also be numbered by the addition of lower caseletters to the position number of the amino acid residue preceding theinserted amino acid residue(s), in this example: *#₁aK*#₁bG.

Multiple alterations: Variants comprising multiple alterations areseparated by addition marks (“+”) or by commas (,), e.g., “Val #₁Trp+Pro#₂Gly” or “V #₁W, P #₂G” representing a substitution of valine andproline at positions #₁ and #₂ with tryptophan and glycine, respectivelyas described above.

Different alterations: Where different alterations can be introduced ata position, the different alterations are separated by a comma, e.g.,“Val #₁Trp, Lys” or V #₁W, K represents a substitution of valine atposition #₁ with tryptophan or lysine. Thus, “Val #₁Trp, Lys+Pro #₂Asp”designates the following variants: “Val #₁Trp+Pro #₂Asp”, “Val #₁Lys+Pro#₂Asp” or V #₁W, K+P #₂D.

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides novel DNases obtained from Aspergillus,in particular, Aspergillus oryzae. The DNases of the invention compriseat least 60% sequence identity to a polypeptide with SEQ ID NO: 1 andcomprise an alteration of at least one amino acid position compared tothe DNase with SEQ ID NO: 1. In one embodiment, the DNase variants havean amino acid sequence comprising at least one substitution of an aminoacid made at a position equivalent to a position in SEQ ID NO: 1. In oneembodiment, the DNase variants have an amino acid sequence comprising atleast one deletion of an amino acid made at a position equivalent to aposition in SEQ ID NO: 1. In one embodiment, the DNase variants have anamino acid sequence comprising at least one insertion of an amino acidmade at a position equivalent to a position in SEQ ID NO: 1. The presentinvention also relates to methods for of generating DNase variants. Theinvention further relates to a screening process comprising the steps ofa) providing a mutant nucleic acid or variant polypeptide therefrom, b)determine a property of interest in the mutant nucleic acid or variantpolypeptide and c) comparing the this property to the same property ofparent nucleic acid or polypeptide (i.e. the nucleic acid or polypeptidenot having the said specific alterations). It will be apparent toskilled artisan that the screening process is not limited to anyspecific property as it depends upon the property determined to bescreened for. A particular preferred screenings method is thehigh-throughput screening, including multiple samples being screenedsimultaneously. Examples of properties which may be screened forincludes wash performance, such as deep-cleaning performance, malodorreduction, high or low pH performance, improved low temperatureperformance, stability such as stability in detergent and/or storagestability. It is not intended that the present invention be limited toany particular method of variant generation or screening. Preferably theDNases of the present invention have at least one improved propertycompared to the parent DNase e.g. compared to SEQ ID NO: 1. Propertiesincludes but are not limited to stability in detergent includingstorage, in wash and thermo stability, wash performance in particulardeep-cleaning performance, increased expression level or malodorreduction.

Embodiments of the invention relates to DNase variants of SEQ ID NO: 1or variants of a DNase having at least 60% identity hereto and tomethods for generating a DNase variant of SEQ ID NO: 1 or a DNase havingat least 60% identity hereto.

One embodiment relates to DNase variants having at least 60% identity toSEQ ID NO: 1, having DNase activity and comprise an alteration at one ormore positions selected from the list consisting of 1, 2, 3, 4, 5, 6, 7,8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62,63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98,99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 185, 186, 187, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 219, 220 and 221, wherein each positioncorresponds to the position of the polypeptide of SEQ ID NO: 1. Thealteration is independently a substitution, insertion or deletion. In anembodiment, the alteration is a substitution. In another embodiment, thealteration is a deletion.

One embodiment of the invention relates to a DNase variant comprising asubstitution at one or more positions selected from the group consistingof 4, 17, 19, 19, 19, 36, 38, 39, 39, 40, 40, 41, 41, 45, 51, 53, 53,54, 55, 57, 64, 64, 64, 64, 64, 64, 66, 67, 67, 68, 68, 68, 68, 69, 69,69, 69, 69, 69, 69, 69, 70, 70, 70, 71, 72, 74, 74, 75, 77, 82, 82, 83,83, 83, 83, 83, 84, 85, 85, 85, 86, 86, 86, 88, 88, 91, 99, 101, 105,105, 105, 105, 106, 115, 116, 135, 136, 138, 138, 138, 138, 139, 140,140, 140, 141, 151, 152, 152, 152, 152, 153, 154, 162, 163, 164, 166,166, 168, 169, 169, 173, 173, 173, 182, 183, 184, 185, 186, 189, 189,212, 212 and 215, wherein each position corresponds to the position ofthe polypeptide of SEQ ID NO: 1.

In some embodiment, the DNase variant has sequence identity of at least60%, e.g., at least 65%, at least 70%, at least 75%, at least 80%, atleast 85%, at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, or atleast 99%, but less than 100%, to the amino acid sequence of the parentDNase.

In another embodiment, the variant has at least 60%, e.g., at least 65%,at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, atleast 91%, at least 92%, at least 93%, at least 94%, at least 95%, suchas at least 96%, at least 97%, at least 98%, or at least 99%, but lessthan 100%, sequence identity to the polypeptide shown in SEQ ID NO: 1.

The percent sequence identity is determined using the using theNeedleman-Wunsch algorithm (Needleman and Wunsch, 1970, J. Mol. Biol.48: 443-453) as described in “Conventions for Designation of Variants”.

The invention further relates to variants of a DNase parent comprisingSEQ ID NO: 1 wherein said variant comprises an alteration compared toSEQ ID NO: 1 in at least one position selected from the positions: 1, 2,3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110,111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124,125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138,139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152,153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166,167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180,181, 182, 183, 185, 186, 187, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 219, 220 and 221 correspondingto the positions of SEQ ID NO: 1, wherein the variant has an amino acidsequence which is at least 60%, at least 70%, at least 80%, at least 90%or at least 95% identical to SEQ ID NO: 1.

The invention further relates to variants of a DNase parent comprisingSEQ ID NO: 1 wherein said variant comprises an alteration compared toSEQ ID NO: 1 in at least one position selected from the positions: 4,17, 19, 19, 19, 36, 38, 39, 39, 40, 40, 41, 41, 45, 51, 53, 53, 54, 55,57, 64, 64, 64, 64, 64, 64, 66, 67, 67, 68, 68, 68, 68, 69, 69, 69, 69,69, 69, 69, 69, 70, 70, 70, 71, 72, 74, 74, 75, 77, 82, 82, 83, 83, 83,83, 83, 84, 85, 85, 85, 86, 86, 86, 88, 88, 91, 99, 101, 105, 105, 105,105, 106, 115, 116, 135, 136, 138, 138, 138, 138, 139, 140, 140, 140,141, 151, 152, 152, 152, 152, 153, 154, 162, 163, 164, 166, 166, 168,169, 169, 173, 173, 173, 182, 183, 184, 185, 186, 189, 189, 212, 212 and215, corresponding to the positions of SEQ ID NO: 1, wherein the varianthas an amino acid sequence which is at least 60%, at least 70%, at least80%, at least 90% or at least 95% identical to SEQ ID NO: 1.

The alteration is independently a substitution, insertion or deletion.In an embodiment, the alteration is a substitution. In anotherembodiment, the alteration is a deletion. In one aspect, the number ofalterations in the variants of the present invention is 1-20, e.g., 1-10and 1-5, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 alterations.

In some preferred embodiments, the DNase variants have at least 60%,such as at least 70%, such as at least 80% such as at least 90% but lessthan 100% sequence identity to SEQ ID NO: 1, have DNase activity andcomprises one or more alterations selected from the group consisting of:V1*, V1A, V1D, V1E, V1F, V1G, V1H, V1I, V1K, V1L, V1M, V1N, V1P, V1Q,V1R, V1S, V1T, V1W, V1Y, P2*, P2A, P2D, P2E, P2F, P2G, P2H, P2I, P2K,P2L, P2M, P2N, P2Q, P2R, P2S, P2T, P2V, P2W, P2Y, V3*, V3A, V3C, V3D,V3E, V3F, V3G, V3H, V3I, V3K, V3L, V3M, V3N, V3P, V3R, V3S, V3T, V3W,V3Y, N4*, N4A, N4D, N4E, N4F, N4G, N4H, N4I, N4K, N4L, N4M, N4P, N4Q,N4R, N4S, N4T, N4V, N4W, N4Y, P5*, P5A, P5D, P5E, P5F, P5G, P5H, P5I,P5K, P5L, P5M, P5N, P5Q, P5R, P5S, P5T, P5V, P5W, P5Y, E6*, E6A, E6D,E6F, E6G, E6H, E6I, E6K, E6L, E6M, E6N, E6P, E6Q, E6R, E6S, E6T, E6V,E6W, E6Y, P7*, P7A, P7D, P7E, P7F, P7G, P7H, P7I, P7K, P7L, P7M, P7N,P7Q, P7R, P7S, P7T, P7V, P7W, P7Y, D8*, D8A, D8E, D8F, D8G, D8H, D8I,D8K, D8L, D8M, D8N, D8P, D8Q, D8R, D8S, D8T, D8V, D8W, D8Y, A9*, A9D,A9E, A9F, A9G, A9H, A9I, A9K, A9L, A9M, A9N, A9P, A9Q, A9R, A9S, A9T,A9V, A9W, A9Y, T10*, T10A, T10D, T10E, T10F, T10G, T10H, T10I, T10K,T10L, T10M, T10N, T10P, T10Q, T10R, T10S, T10V, T10W, T10Y, S11*, S11A,S11D, S11E, S11F, S11G, S11H, S11I, S11K, S11L, S11M, S11N, S11P, S11Q,S11R, S11T, S11V, S11W, S11Y, V12*, V12A, V12D, V12E, V12F, V12G, V12H,V12I, V12K, V12L, V12M, V12N, V12P, V12Q, V12R, V12S, V12T, V12W, V12Y,E13*, E13A, E13D, E13F, E13G, E13H, E13I, E13K, E13L, E13M, E13N, E13P,E13Q, E13R, E13S, E13T, E13V, E13W, E13Y, N14*, N14A, N14D, N14E, N14F,N14G, N14H, N14I, N14K, N14L, N14M, N14P, N14Q, N14R, N14S, N14T, N14V,N14W, N14Y, V15*, V15A, V15D, V15E, V15F, V15G, V15H, V15I, V15K, V15L,V15M, V15N, V15P, V15Q, V15R, V15S, V15T, V15W, V15Y, A16*, A16D, A16E,A16F, A16G, A16H, A16I, A16K, A16L, A16M, A16N, A16P, A16Q, A16R, A16S,A16T, A16V, A16W, A16Y, L17*, L17A, L17D, L17E, L17F, L17G, L17H, L17I,L17K, L17M, L17N, L17P, L17Q, L17R, L17S, L17T, L17V, L17W, L17Y, K18*,K18A, K18D, K18E, K18F, K18G, K18H, K18I, K18L, K18M, K18N, K18P, K18Q,K18R, K18S, K18T, K18V, K18W, K18Y, T19*, T19A, T19D, T19E, T19F, T19G,T19H, T19I, T19K, T19L, T19M, T19N, T19P, T19Q, T19R, T19S, T19V, T19W,T19Y, G20*, G20A, G20D, G20E, G20F, G20H, G20I, G20K, G20L, G20M, G20N,G20P, G20Q, G20R, G20S, G20T, G20V, G20W, G20Y, S21*, S21A, S21D, S21E,S21F, S21G, S21H, S21I, S21K, S21L, S21M, S21N, S21P, S21Q, S21R, S21T,S21V, S21W, S21Y, G22*, G22A, G22D, G22E, G22F, G22H, G22I, G22K, G22L,G22M, G22N, G22P, G22Q, G22R, G22S, G22T, G22V, G22W, G22Y, D23*, D23A,D23E, D23F, D23G, D23H, D23I, D23K, D23L, D23M, D23N, D23P, D23Q, D23R,D23S, D23T, D23V, D23W, D23Y, S24*, S24A, S24D, S24E, S24F, S24G, S24H,S24I, S24K, S24L, S24M, S24N, S24P, S24Q, S24R, S24T, S24V, S24W, S24Y,Q25*, Q25A, Q25D, Q25E, Q25F, Q25G, Q25H, Q25I, Q25K, Q25L, Q25M, Q25N,Q25P, Q25R, Q25S, Q25T, Q25V, Q25W, Q25Y, S26*, S26A, S26D, S26E, S26F,S26G, S26H, S26I, S26K, S26L, S26M, S26N, S26P, S26Q, S26R, S26T, S26V,S26W, S26Y, D27*, D27A, D27E, D27F, D27G, D27H, D27I, D27K, D27L, D27M,D27N, D27P, D27Q, D27R, D27S, D27T, D27V, D27W, D27Y, P28*, P28A, P28D,P28E, P28F, P28G, P28H, P28I, P28K, P28L, P28M, P28N, P28Q, P28R, P28S,P28T, P28V, P28W, P28Y, I29*, I29A, I29D, I29E, I29F, I29G, I29H, I29K,I29L, I29M, I29N, I29P, I29Q, I29R, I29S, I29T, I29V, I29W, I29Y, K30*,K30A, K30D, K30E, K30F, K30G, K30H, K30I, K30L, K30M, K30N, K30P, K30Q,K30R, K30S, K30T, K30V, K30W, K30Y, A31*, A31D, A31E, A31F, A31G, A31H,A31I, A31K, A31L, A31M, A31N, A31P, A31Q, A31R, A31S, A31T, A31V, A31W,A31Y, D32*, D32A, D32E, D32F, D32G, D32H, D32I, D32K, D32L, D32M, D32N,D32P, D32Q, D32R, D32S, D32T, D32V, D32W, D32Y, L33*, L33A, L33D, L33E,L33F, L33G, L33H, L33I, L33K, L33M, L33N, L33P, L33Q, L33R, L33S, L33T,L33V, L33W, L33Y, E34*, E34A, E34D, E34F, E34G, E34H, E34I, E34K, E34L,E34M, E34N, E34P, E34Q, E34R, E34S, E34T, E34V, E34W, E34Y, V35*, V35A,V35D, V35E, V35F, V35G, V35H, V35I, V35K, V35L, V35M, V35N, V35P, V35Q,V35R, V35S, V35T, V35W, V35Y, K36*, K36A, K36D, K36E, K36F, K36G, K36H,K36I, K36L, K36M, K36N, K36P, K36Q, K36R, K36S, K36T, K36V, K36W, K36Y,G37*, G37A, G37D, G37E, G37F, G37H, G37I, G37K, G37L, G37M, G37N, G37P,G37Q, G37R, G37S, G37T, G37V, G37W, G37Y, Q38*, Q38A, Q38D, Q38E, Q38F,Q38G, Q38H, Q38I, Q38K, Q38L, Q38M, Q38N, Q38P, Q38R, Q38S, Q38T, Q38V,Q38W, Q38Y, S39*, S39A, S39D, S39E, S39F, S39G, S39H, S39I, S39K, S39L,S39M, S39N, S39P, S39Q, S39R, S39T, S39V, S39W, S39Y, A40*, A40D, A40E,A40F, A40G, A40H, A40I, A40K, A40L, A40M, A40N, A40P, A40Q, A40R, A40S,A40T, A40V, A40W, A40Y, L41*, L41A, L41D, L41E, L41F, L41G, L41H, L41I,L41K, L41M, L41N, L41P, L41Q, L41R, L41S, L41T, L41V, L41W, L41Y, P42*,P42A, P42D, P42E, P42F, P42G, P42H, P42I, P42K, P42L, P42M, P42N, P42Q,P42R, P42S, P42T, P42V, P42W, P42Y, F43*, F43A, F43D, F43E, F43G, F43H,F43I, F43K, F43L, F43M, F43N, F43P, F43Q, F43R, F43S, F43T, F43V, F43W,F43Y, D44*, D44A, D44E, D44F, D44G, D44H, D44I, D44K, D44L, D44M, D44N,D44P, D44Q, D44R, D44S, D44T, D44V, D44W, D44Y, V45*, V45A, V45D, V45E,V45F, V45G, V45H, V45I, V45K, V45L, V45M, V45N, V45P, V45Q, V45R, V45S,V45T, V45W, V45Y, D46*, D46A, D46E, D46F, D46G, D46H, D46I, D46K, D46L,D46M, D46N, D46P, D46Q, D46R, D46S, D46T, D46V, D46W, D46Y, C47A, C47D,C47E, C47F, C47G, C47H, C47I, C47K, C47L, C47M, C47N, C47P, C47Q, C47R,C47S, C47T, C47V, C47W, C47Y, W48*, W48A, W48D, W48E, W48F, W48G, W48H,W48I, W48K, W48L, W48M, W48N, W48P, W48Q, W48R, W48S, W48T, W48V, W48Y,A49*, A49D, A49E, A49F, A49G, A49H, A49I, A49K, A49L, A49M, A49N, A49P,A49Q, A49R, A49S, A49T, A49V, A49W, A49Y, I50*, I50A, I50D, I50E, I50F,I50G, I50H, I50K, I50L, I50M, I50N, I50P, I50Q, I50R, I50S, I50T, I50V,I50W, 150Y, L51*, L51A, L51D, L51E, L51F, L51G, L51H, L51I, L51K, L51M,L51N, L51P, L51Q, L51R, L51S, L51T, L51V, L51W, L51Y, K53*, K53A, K53D,K53E, K53F, K53G, K53H, K53I, K53L, K53M, K53N, K53P, K53Q, K53R, K53S,K53T, K53V, K53W, K53Y, G54*, G54A, G54D, G54E, G54F, G54H, G54I, G54K,G54L, G54M, G54N, G54P, G54Q, G54R, G54S, G54T, G54V, G54W, G54Y, A55*,A55D, A55E, A55F, A55G, A55H, A55I, A55K, A55L, A55M, A55N, A55P, A55Q,A55R, A55S, A55T, A55V, A55W, A55Y, P56*, P56A, P56D, P56E, P56F, P56G,P56H, P56I, P56K, P56L, P56M, P56N, P56Q, P56R, P56S, P56T, P56V, P56W,P56Y, N57*, N57A, N57D, N57E, N57F, N57G, N57H, N57I, N57K, N57L, N57M,N57P, N57Q, N57R, N57S, N57T, N57V, N57W, N57Y, V58*, V58A, V58D, V58E,V58F, V58G, V58H, V58I, V58K, V58L, V58M, V58N, V58P, V58Q, V58R, V58S,V58T, V58W, V58YL59*, L59A, L59D, L59E, L59F, L59G, L59H, L59I, L59K,L59M, L59N, L59P, L59Q, L59R, L59S, L59T, L59V, L59W, L59Y, Q60*, Q60A,Q60D, Q60E, Q60F, Q60G, Q60H, Q60I, Q60K, Q60L, Q60M, Q60N, Q60P, Q60R,Q60S, Q60T, Q60V, Q60W, Q60Y, R61*, R61A, R61D, R61E, R61F, R61G, R61H,R61I, R61K, R61L, R61M, R61N, R61P, R61Q, R61S, R61T, R61V, R61W, R61Y,V62*, V62A, V62D, V62E, V62F, V62G, V62H, V62I, V62K, V62L, V62M, V62N,V62P, V62Q, V62R, V62S, V62T, V62W, V62Y, N63*, N63A, N63D, N63E, N63F,N63G, N63H, N63I, N63K, N63L, N63M, N63P, N63Q, N63R, N63S, N63T, N63V,N63W, N63Y, E64*, E64A, E64D, E64F, E64G, E64H, E64I, E64K, E64L, E64M,E64N, E64P, E64Q, E64R, E64S, E64T, E64V, E64W, E64Y, K65*, K65A, K65D,K65E, K65F, K65G, K65H, K65I, K65L, K65M, K65N, K65P, K65Q, K65R, K65S,K65T, K65V, K65W, K65Y, T66*, T66A, T66D, T66E, T66F, T66G, T66H, T66I,T66K, T66L, T66M, T66N, T66P, T66Q, T66R, T66S, T66V, T66W, T66Y, K67*,K67A, K67D, K67E, K67F, K67G, K67H, K67I, K67L, K67M, K67N, K67P, K67Q,K67R, K67S, K67T, K67V, K67W, K67Y, N68*, N68A, N68D, N68E, N68F, N68G,N68H, N68I, N68K, N68L, N68M, N68P, N68Q, N68R, N68S, N68T, N68V, N68W,N68Y, S69*, S69A, S69D, S69E, S69F, S69G, S69H, S69I, S69K, S69L, S69M,S69N, S69P, S69Q, S69R, S69T, S69V, S69W, S69Y, N70*, N70A, N70D, N70E,N70F, N70G, N70H, N70I, N70K, N70L, N70M, N70P, N70Q, N70R, N70S, N70T,N70V, N70W, N70Y, R71*, R71A, R71D, R71E, R71F, R71G, R71H, R71I, R71K,R71L, R71M, R71N, R71P, R71Q, R71S, R71T, R71V, R71W, R71Y, D72*, D72A,D72E, D72F, D72G, D72H, D72I, D72K, D72L, D72M, D72N, D72P, D72Q, D72R,D72S, D72T, D72V, D72W, D72Y, R73*, R73A, R73D, R73E, R73F, R73G, R73H,R73I, R73K, R73L, R73M, R73N, R73P, R73Q, R73S, R73T, R73V, R73W, R73Y,S74*, S74A, S74D, S74E, S74F, S74G, S74H, S74I, S74K, S74L, S74M, S74N,S74P, S74Q, S74R, S74T, S74V, S74W, S74Y, G75*, G75A, G75D, G75E, G75F,G75H, G75I, G75K, G75L, G75M, G75N, G75P, G75Q, G75R, G75S, G75T, G75V,G75W, G75Y, A76*, A76D, A76E, A76F, A76G, A76H, A76I, A76K, A76L, A76M,A76N, A76P, A76Q, A76R, A76S, A76T, A76V, A76W, A76Y, N77*, N77A, N77D,N77E, N77F, N77G, N77H, N77I, N77K, N77L, N77M, N77P, N77Q, N77R, N77S,N77T, N77V, N77W, N77Y, K78*, K78A, K78D, K78E, K78F, K78G, K78H, K78I,K78L, K78M, K78N, K78P, K78Q, K78R, K78S, K78T, K78V, K78W, K78Y, G79*,G79A, G79D, G79E, G79F, G79H, G79I, G79K, G79L, G79M, G79N, G79P, G79Q,G79R, G79S, G79T, G79V, G79W, G79Y, P80*, P80A, P80D, P80E, P80F, P80G,P80H, P80I, P80K, P80L, P80M, P80N, P80Q, P80R, P80S, P80T, P80V, P80W,P80Y, F81*, F81A, F81D, F81E, F81G, F81H, F81I, F81K, F81L, F81M, F81N,F81P, F81Q, F81R, F81S, F81T, F81V, F81W, F81Y, K82*, K82A, K82D, K82E,K82F, K82G, K82H, K82I, K82L, K82M, K82N, K82P, K82Q, K82R, K82S, K82T,K82V, K82W, K82Y, D83*, D83A, D83E, D83F, D83G, D83H, D83I, D83K, D83L,D83M, D83N, D83P, D83Q, D83R, D83S, D83T, D83V, D83W, D83Y, P84*, P84A,P84D, P84E, P84F, P84G, P84H, P84I, P84K, P84L, P84M, P84N, P84Q, P84R,P84S, P84T, P84V, P84W, P84Y, Q85*, Q85A, Q85D, Q85E, Q85F, Q85G, Q85H,Q85I, Q85K, Q85L, Q85M, Q85N, Q85P, Q85R, Q85S, Q85T, Q85V, Q85W, Q85Y,K86*, K86A, K86D, K86E, K86F, K86G, K86H, K86I, K86L, K86M, K86N, K86P,K86Q, K86R, K86S, K86T, K86V, K86W, K86Y, W87*, W87A, W87D, W87E, W87F,W87G, W87H, W87I, W87K, W87L, W87M, W87N, W87P, W87Q, W87R, W87S, W87T,W87V, W87Y, G88*, G88A, G88D, G88E, G88F, G88H, G88I, G88K, G88L, G88M,G88N, G88P, G88Q, G88R, G88S, G88T, G88V, G88W, G88Y, I89*, I89A, I89D,I89E, I89F, I89G, I89H, I89K, I89L, I89M, I89N, I89P, I89Q, I89R, I89S,I89T, I89V, I89W, I89Y, K90*, K90A, K90D, K90E, K90F, K90G, K90H, K90I,K90L, K90M, K90N, K90P, K90Q, K90R, K90S, K90T, K90V, K90W, K90Y, A91*,A91D, A91E, A91F, A91G, A91H, A91I, A91K, A91L, A91M, A91N, A91P, A91Q,A91R, A91S, A91T, A91V, A91W, A91Y, L92*, L92A, L92D, L92E, L92F, L92G,L92H, L92I, L92K, L92M, L92N, L92P, L92Q, L92R, L92S, L92T, L92V, L92W,L92Y, P93*, P93A, P93D, P93E, P93F, P93G, P93H, P93I, P93K, P93L, P93M,P93N, P93Q, P93R, P93S, P93T, P93V, P93W, P93Y, P94*, P94A, P94D, P94E,P94F, P94G, P94H, P94I, P94K, P94L, P94M, P94N, P94Q, P94R, P94S, P94T,P94V, P94W, P94Y, K95*, K95A, K95D, K95E, K95F, K95G, K95H, K95I, K95L,K95M, K95N, K95P, K95Q, K95R, K95S, K95T, K95V, K95W, K95Y, N96*, N96A,N96D, N96E, N96F, N96G, N96H, N96I, N96K, N96L, N96M, N96P, N96Q, N96R,N96S, N96T, N96V, N96W, N96Y, P97*, P97A, P97D, P97E, P97F, P97G, P97H,P97I, P97K, P97L, P97M, P97N, P97Q, P97R, P97S, P97T, P97V, P97W, P97Y,S98*, S98A, S98D, S98E, S98F, S98G, S98H, S98I, S98K, S98L, S98M, S98N,S98P, S98Q, S98R, S98T, S98V, S98W, S98Y, W99*, W99A, W99D, W99E, W99F,W99G, W99H, W99I, W99K, W99L, W99M, W99N, W99P, W99Q, W99R, W99S, W99T,W99V, W99Y, S100*, S100A, S100D, S100E, S100F, S100G, S100H, S100I,S100K, S100L, S100M, S100N, S100P, S100Q, S100R, S100T, S100V, S100W,S100Y, A101*, A101D, A101E, A101F, A101G, A101H, A101I, A101K, A101L,A101M, A101N, A101P, A101Q, A101R, A101S, A101T, A101V, A101W, A101Y,Q102*, Q102A, Q102D, Q102E, Q102F, Q102G, Q102H, Q102I, Q102K, Q102L,Q102M, Q102N, Q102P, Q102R, Q102S, Q102T, Q102V, Q102W, Q102Y, D103*,D103A, D103E, D103F, D103G, D103H, D103I, D103K, D103L, D103M, D103N,D103P, D103Q, D103R, D103S, D103T, D103V, D103W, D103Y, F104*, F104A,F104D, F104E, F104G, F104H, F104I, F104K, F104L, F104M, F104N, F104P,F104Q, F104R, F104S, F104T, F104V, F104W, F104Y, K105*, K105A, K105D,K105E, K105F, K105G, K105H, K105I, K105L, K105M, K105N, K105P, K105Q,K105R, K105S, K105T, K105V, K105W, K105Y, S106*, S106A, S106D, S106E,S106F, S106G, S106H, S106I, S106K, S106L, S106M, S106N, S106P, S106Q,S106R, S106T, S106V, S106W, S106Y, P107*, P107A, P107D, P107E, P107F,P107G, P107H, P107I, P107K, P107L, P107M, P107N, P107Q, P107R, P107S,P107T, P107V, P107W, P107Y, E108*, E108A, E108D, E108F, E108G, E108H,E108I, E108K, E108L, E108M, E108N, E108P, E108Q, E108R, E108S, E108T,E108V, E108W, E108Y, E109*, E109A, E109D, E109F, E109G, E109H, E109I,E109K, E109L, E109M, E109N, E109P, E109Q, E109R, E109S, E109T, E109V,E109W, E109Y, Y110*, Y110A, Y110D, Y110E, Y110F, Y110G, Y110H, Y110I,Y110K, Y110L, Y110M, Y110N, Y110P, Y110Q, Y110R, Y110S, Y110T, Y110V,Y110W, A111*, A111D, A111E, A111F, A111G, A111H, A111I, A111K, A111L,A111M, A111N, A111P, A111Q, A111R, A111S, A111T, A111V, A111W, A111Y,F112*, F112A, F112D, F112E, F112G, F112H, F112I, F112K, F112L, F112M,F112N, F112P, F112Q, F112R, F112S, F112T, F112V, F112W, F112Y, A113*,A113D, A113E, A113F, A113G, A113H, A113I, A113K, A113L, A113M, A113N,A113P, A113Q, A113R, A113S, A113T, A113V, A113W, A113Y, S114*, S114A,S114D, S114E, S114F, S114G, S114H, S114I, S114K, S114L, S114M, S114N,S114P, S114Q, S114R, S114T, S114V, S114W, S114Y, S115*, S115A, S115D,S115E, S115F, S115G, S115H, S115I, S115K, S115L, S115M, S115N, S115P,S115Q, S115R, S115T, S115V, S115W, S115Y, L116*, L116A, L116D, L116E,L116F, L116G, L116H, L116I, L116K, L116M, L116N, L116P, L116Q, L116R,L116S, L116T, L116V, L116W, L116Y, Q117*, Q117A, Q117D, Q117E, Q117F,Q117G, Q117H, Q117I, Q117K, Q117L, Q117M, Q117N, Q117P, Q117R, Q117S,Q117T, Q117V, Q117W, Q117Y, G118*, G118A, G118D, G118E, G118F, G118H,G118I, G118K, G118L, G118M, G118N, G118P, G118Q, G118R, G118S, G118T,G118V, G118W, G118Y, G119*, G119A, G119D, G119E, G119F, G119H, G119I,G119K, G119L, G119M, G119N, G119P, G119Q, G119R, G119S, G119T, G119V,G119W, G119Y, T120*, T120A, T120D, T120E, T120F, T120G, T120H, T120I,T120K, T120L, T120M, T120N, T120P, T120Q, T120R, T120S, T120V, T120W,T120Y, N121*, N121A, N121D, N121E, N121F, N121G, N121H, N121I, N121K,N121L, N121M, N121P, N121Q, N121R, N121S, N121T, N121V, N121W, N121Y,A122*, A122D, A122E, A122F, A122G, A122H, A122I, A122K, A122L, A122M,A122N, A122P, A122Q, A122R, A122S, A122T, A122V, A122W, A122Y, I123*,I123A, I123D, I123E, I123F, I123G, I123H, I123K, I123L, I123M, I123N,I123P, I123Q, I123R, I123S, I123T, I123V, I123W, I123Y, L124*, L124A,L124D, L124E, L124F, L124G, L124H, L124I, L124K, L124M, L124N, L124P,L124Q, L124R, L124S, L124T, L124V, L124W, L124Y, A125*, A125D, A125E,A125F, A125G, A125H, A125I, A125K, A125L, A125M, A125N, A125P, A125Q,A125R, A125S, A125T, A125V, A125W, A125Y, P126*, P126A, P126D, P126E,P126F, P126G, P126H, P126I, P126K, P126L, P126M, P126N, P126Q, P126R,P126S, P126T, P126V, P126W, P126Y, V127*, V127A, V127D, V127E, V127F,V127G, V127H, V127I, V127K, V127L, V127M, V127N, V127P, V127Q, V127R,V127S, V127T, V127W, V127Y, N128*, N128A, N128D, N128E, N128F, N128G,N128H, N128I, N128K, N128L, N128M, N128P, N128Q, N128R, N128S, N128T,N128V, N128W, N128Y, L129*, L129A, L129D, L129E, L129F, L129G, L129H,L129I, L129K, L129M, L129N, L129P, L129Q, L129R, L129S, L129T, L129V,L129W, L129Y, A130*, A130D, A130E, A130F, A130G, A130H, A130I, A130K,A130L, A130M, A130N, A130P, A130Q, A130R, A130S, A130T, A130V, A130W,A130Y, S131*, S131A, S131D, S131E, S131F, S131G, S131H, S131I, S131K,S131L, S131M, S131N, S131P, S131Q, S131R, S131T, S131V, S131W, S131Y,Q132*, Q132A, Q132D, Q132E, Q132F, Q132G, Q132H, Q132I, Q132K, Q132L,Q132M, Q132N, Q132P, Q132R, Q132S, Q132T, Q132V, Q132W, Q132Y, N133*,N133A, N133D, N133E, N133F, N133G, N133H, N133I, N133K, N133L, N133M,N133P, N133Q, N133R, N133S, N133T, N133V, N133W, N133Y, S134*, S134A,S134D, S134E, S134F, S134G, S134H, S134I, S134K, S134L, S134M, S134N,S134P, S134Q, S134R, S134T, S134V, S134W, S134Y, Q135*, Q135A, Q135D,Q135E, Q135F, Q135G, Q135H, Q135I, Q135K, Q135L, Q135M, Q135N, Q135P,Q135R, Q135S, Q135T, Q135V, Q135W, Q135Y, G136*, G136A, G136D, G136E,G136F, G136H, G136I, G136K, G136L, G136M, G136N, G136P, G136Q, G136R,G136S, G136T, G136V, G136W, G136Y, G137*, G137A, G137D, G137E, G137F,G137H, G137I, G137K, G137L, G137M, G137N, G137P, G137Q, G137R, G137S,G137T, G137V, G137W, G137Y, V138*, V138A, V138D, V138E, V138F, V138G,V138H, V138I, V138K, V138L, V138M, V138N, V138P, V138Q, V138R, V138S,V138T, V138W, V138Y, L139*, L139A, L139D, L139E, L139F, L139G, L139H,L139I, L139K, L139M, L139N, L139P, L139Q, L139R, L139S, L139T, L139V,L139W, L139Y, N140*, N140A, N140D, N140E, N140F, N140G, N140H, N140I,N140K, N140L, N140M, N140P, N140Q, N140R, N140S, N140T, N140V, N140W,N140Y, G141*, G141A, G141D, G141E, G141F, G141H, G141I, G141K, G141L,G141M, G141N, G141P, G141Q, G141R, G141S, G141T, G141V, G141W, G141Y,F142*, F142A, F142D, F142E, F142G, F142H, F142I, F142K, F142L, F142M,F142N, F142P, F142Q, F142R, F142S, F142T, F142V, F142W, F142Y, Y143*,Y143A, Y143D, Y143E, Y143F, Y143G, Y143H, Y143I, Y143K, Y143L, Y143M,Y143N, Y143P, Y143Q, Y143R, Y143S, Y143T, Y143V, Y143W, S144*, S144A,S144D, S144E, S144F, S144G, S144H, S144I, S144K, S144L, S144M, S144N,S144P, S144Q, S144R, S144T, S144V, S144W, S144Y, A145*, A145D, A145E,A145F, A145G, A145H, A145I, A145K, A145L, A145M, A145N, A145P, A145Q,A145R, A145S, A145T, A145V, A145W, A145Y, N146*, N146A, N146D, N146E,N146F, N146G, N146H, N146I, N146K, N146L, N146M, N146P, N146Q, N146R,N146S, N146T, N146V, N146W, N146Y, K147*, K147A, K147D, K147E, K147F,K147G, K147H, K147I, K147L, K147M, K147N, K147P, K147Q, K147R, K147S,K147T, K147V, K147W, K147Y, V148*, V148A, V148D, V148E, V148F, V148G,V148H, V148I, V148K, V148L, V148M, V148N, V148P, V148Q, V148R, V148S,V148T, V148W, V148Y, A149*, A149D, A149E, A149F, A149G, A149H, A149I,A149K, A149L, A149M, A149N, A149P, A149Q, A149R, A149S, A149T, A149V,A149W, A149Y, Q150*, Q150A, Q150D, Q150E, Q150F, Q150G, Q150H, Q150I,Q150K, Q150L, Q150M, Q150N, Q150P, Q150R, Q150S, Q150T, Q150V, Q150W,Q150Y, F151*, F151A, F151D, F151E, F151G, F151H, F151I, F151K, F151L,F151M, F151N, F151P, F151Q, F151R, F151S, F151T, F151V, F151W, F151Y,D152*, D152A, D152E, D152F, D152G, D152H, D152I, D152K, D152L, D152M,D152N, D152P, D152Q, D152R, D152S, D152T, D152V, D152W, D152Y, P153*,P153A, P153D, P153E, P153F, P153G, P153H, P153I, P153K, P153L, P153M,P153N, P153Q, P153R, P153S, P153T, P153V, P153W, P153Y, S154*, S154A,S154D, S154E, S154F, S154G, S154H, S154I, S154K, S154L, S154M, S154N,S154P, S154Q, S154R, S154T, S154V, S154W, S154Y, K155*, K155A, K155D,K155E, K155F, K155G, K155H, K155I, K155L, K155M, K155N, K155P, K155Q,K155R, K155S, K155T, K155V, K155W, K155Y, P156*, P156A, P156D, P156E,P156F, P156G, P156H, P156I, P156K, P156L, P156M, P156N, P156Q, P156R,P156S, P156T, P156V, P156W, P156Y, Q157*, Q157A, Q157D, Q157E, Q157F,Q157G, Q157H, Q157I, Q157K, Q157L, Q157M, Q157N, Q157P, Q157R, Q157S,Q157T, Q157V, Q157W, Q157Y, Q158*, Q158A, Q158D, Q158E, Q158F, Q158G,Q158H, Q158I, Q158K, Q158L, Q158M, Q158N, Q158P, Q158R, Q158S, Q158T,Q158V, Q158W, Q158Y, T159*, T159A, T159D, T159E, T159F, T159G, T159H,T159I, T159K, T159L, T159M, T159N, T159P, T159Q, T159R, T159S, T159V,T159W, T159Y, K160*, K160A, K160D, K160E, K160F, K160G, K160H, K160I,K160L, K160M, K160N, K160P, K160Q, K160R, K160S, K160T, K160V, K160W,K160Y, G161*, G161A, G161D, G161E, G161F, G161H, G161I, G161K, G161L,G161M, G161N, G161P, G161Q, G161R, G161S, G161T, G161V, G161W, G161Y,T162A, T162D, T162E, T162F, T162G, T162H, T162I, T162K, T162L, T162M,T162N, T162P, T162Q, T162R, T162S, T162T, T162V, T162W, T162Y, W163*,W163A, W163D, W163E, W163F, W163G, W163H, W163I, W163K, W163L, W163M,W163N, W163P, W163Q, W163R, W163S, W163T, W163V, W163Y, F164*, F164A,F164D, F164E, F164G, F164H, F164I, F164K, F164L, F164M, F164N, F164P,F164Q, F164R, F164S, F164T, F164V, F164W, F164Y, Q165*, Q165A, Q165D,Q165E, Q165F, Q165G, Q165H, Q165I, Q165K, Q165L, Q165M, Q165N, Q165P,Q165R, Q165S, Q165T, Q165V, Q165W, Q165Y, I166*, I166A, I166D, I166E,I166F, I166G, I166H, I166K, I166L, I166M, I166N, I166P, I166Q, I166R,I166S, I166T, I166V, I166W, I166Y, T167*, T167A, T167D, T167E, T167F,T167G, T167H, T167I, T167K, T167L, T167M, T167N, T167P, T167Q, T167R,T167S, T167V, T167W, T167Y, K168*, K168A, K168D, K168E, K168F, K168G,K168H, K168I, K168L, K168M, K168N, K168P, K168Q, K168R, K168S, K168T,K168V, K168W, K168Y, F169*, F169A, F169D, F169E, F169G, F169H, F169I,F169K, F169L, F169M, F169N, F169P, F169Q, F169R, F169S, F169T, F169V,F169W, F169Y, T170*, T170A, T170D, T170E, T170F, T170G, T170H, T170I,T170K, T170L, T170M, T170N, T170P, T170Q, T170R, T170S, T170V, T170W,T170Y, G171*, G171A, G171D, G171E, G171F, G171H, G171I, G171K, G171L,G171M, G171N, G171P, G171Q, G171R, G171S, G171T, G171V, G171W, G171Y,A172*, A1720, A172E, A172F, A172G, A172H, A172I, A172K, A172L, A172M,A172N, A172P, A172Q, A172R, A172S, A172T, A172V, A172W, A172Y, A173*,A173D, A173E, A173F, A173G, A173H, A173I, A173K, A173L, A173M, A173N,A173P, A173Q, A173R, A173S, A173T, A173V, A173W, A173Y, G174*, G174A,G174D, G174E, G174F, G174H, G174I, G174K, G174L, G174M, G174N, G174P,G174Q, G174R, G174S, G174T, G174V, G174W, G174Y, P175*, P175A, P175D,P175E, P175F, P175G, P175H, P175I, P175K, P175L, P175M, P175N, P175Q,P175R, P175S, P175T, P175V, P175W, P175Y, Y176*, Y176A, Y176D, Y176E,Y176F, Y176G, Y176H, Y176I, Y176K, Y176L, Y176M, Y176N, Y176P, Y176Q,Y176R, Y176S, Y176T, Y176V, Y176W, K178*, K178A, K178D, K178E, K178F,K178G, K178H, K178I, K178L, K178M, K178N, K178P, K178Q, K178R, K178S,K178T, K178V, K178W, K178Y, A179*, A179D, A179E, A179F, A179G, A179H,A179I, A179K, A179L, A179M, A179N, A179P, A179Q, A179R, A179S, A179T,A179V, A179W, A179Y, L180*, L180A, L180D, L180E, L180F, L180G, L180H,L180I, L180K, L180M, L180N, L180P, L180Q, L180R, L180S, L180T, L180V,L180W, L180Y, G181*, G181A, G181D, G181E, G181F, G181H, G181I, G181K,G181L, G181M, G181N, G181P, G181Q, G181R, G181S, G181T, G181V, G181W,G181Y, S182*, S182A, S182D, S182E, S182F, S182G, S182H, S182I, S182K,S182L, S182M, S182N, S182P, S182Q, S182R, S182T, S182V, S182W, S182Y,N183*, N183A, N183D, N183E, N183F, N183G, N183H, N183I, N183K, N183L,N183M, N183P, N183Q, N183R, N183S, N183T, N183V, N183W, N183Y, D184*,D184A, D184E, D184F, D184G, D184H, D184I, D184K, D184L, D184M, D184N,D184P, D184Q, D184R, D184S, D184T, D184V, D184W, D184Y, K185*, K185A,K185D, K185E, K185F, K185G, K185H, K185I, K185L, K185M, K185N, K185P,K185Q, K185R, K185S, K185T, K185V, K185W, K185Y, S186*, S186A, S186D,S186E, S186F, S186G, S186H, S186I, S186K, S186L, S186M, S186N, S186P,S186Q, S186R, S186T, S186V, S186W, S186Y, V187*, V187A, V187D, V187E,V187F, V187G, V187H, V187I, V187K, V187L, V187M, V187N, V187P, V187Q,V187R, V187S, V187T, V187W, V187Y, D189*, D189A, D189E, D189F, D189G,D189H, D189I, D189K, D189L, D189M, D189N, D189P, D189Q, D189R, D189S,D189T, D189V, D189W, D189Y, K190*, K190A, K190D, K190E, K190F, K190G,K190H, K190I, K190L, K190M, K190N, K190P, K190Q, K190R, K190S, K190T,K190V, K190W, K190Y, N191*, N191A, N191D, N191E, N191F, N191G, N191H,N191I, N191K, N191L, N191M, N191P, N191Q, N191R, N191S, N191T, N191V,N191W, N191Y, K192*, K192A, K192D, K192E, K192F, K192G, K192H, K192I,K192L, K192M, K192N, K192P, K192Q, K192R, K192S, K192T, K192V, K192W,K192Y, N193*, N193A, N193D, N193E, N193F, N193G, N193H, N193I, N193K,N193L, N193M, N193P, N193Q, N193R, N193S, N193T, N193V, N193W, N193Y,I194*, I194A, I194D, I194E, I194F, I194G, I194H, I194K, I194L, I194M,I194N, I194P, I194Q, I194R, I194S, I194T, I194V, I194W, I194Y, A195*,A195D, A195E, A195F, A195G, A195H, A195I, A195K, A195L, A195M, A195N,A195P, A195Q, A195R, A195S, A195T, A195V, A195W, A195Y, G196*, G196A,G196D, G196E, G196F, G196H, G196I, G196K, G196L, G196M, G196N, G196P,G196Q, G196R, G196S, G196T, G196V, G196W, G196Y, D197*, D197A, D197E,D197F, D197G, D197H, D197I, D197K, D197L, D197M, D197N, D197P, D197Q,D197R, D197S, D197T, D197V, D197W, D197Y, W198*, W198A, W198D, W198E,W198F, W198G, W198H, W198I, W198K, W198L, W198M, W198N, W198P, W198Q,W198R, W198S, W198T, W198V, W198Y, G199*, G199A, G199D, G199E, G199F,G199H, G199I, G199K, G199L, G199M, G199N, G199P, G199Q, G199R, G199S,G199T, G199V, G199W, G199Y, F200*, F200A, F200D, F200E, F200G, F200H,F200I, F200K, F200L, F200M, F200N, F200P, F200Q, F200R, F200S, F200T,F200V, F200W, F200Y, D201*, D201A, D201E, D201F, D201G, D201H, D201I,D201K, D201L, D201M, D201N, D201P, D201Q, D201R, D201S, D201T, D201V,D201W, D201Y, P202*, P202A, P202D, P202E, P202F, P202G, P202H, P202I,P202K, P202L, P202M, P202N, P202Q, P202R, P202S, P202T, P202V, P202W,P202Y, A203*, A203D, A203E, A203F, A203G, A203H, A203I, A203K, A203L,A203M, A203N, A203P, A203Q, A203R, A203S, A203T, A203V, A203W, A203Y,K204*, K204A, K204D, K204E, K204F, K204G, K204H, K204I, K204L, K204M,K204N, K204P, K204Q, K204R, K204S, K204T, K204V, K204W, K204Y, W205*,W205A, W205D, W205E, W205F, W205G, W205H, W205I, W205K, W205L, W205M,W205N, W205P, W205Q, W205R, W205S, W205T, W205V, W205Y, A206*, A206D,A206E, A206F, A206G, A206H, A206I, A206K, A206L, A206M, A206N, A206P,A206Q, A206R, A206S, A206T, A206V, A206W, A206Y, Y207*, Y207A, Y207D,Y207E, Y207F, Y207G, Y207H, Y207I, Y207K, Y207L, Y207M, Y207N, Y207P,Y207Q, Y207R, Y207S, Y207T, Y207V, Y207W, Q208*, Q208A, Q208D, Q208E,Q208F, Q208G, Q208H, Q208I, Q208K, Q208L, Q208M, Q208N, Q208P, Q208R,Q208S, Q208T, Q208V, Q208W, Q208Y, Y209*, Y209A, Y209D, Y209E, Y209F,Y209G, Y209H, Y209I, Y209K, Y209L, Y209M, Y209N, Y209P, Y209Q, Y209R,Y209S, Y209T, Y209V, Y209W, D210*, D210A, D210E, D210F, D210G, D210H,D210I, D210K, D210L, D210M, D210N, D210P, D210Q, D210R, D210S, D210T,D210V, D210W, D210Y, E211*, E211A, E211D, E211F, E211G, E211H, E211I,E211K, E211L, E211M, E211N, E211P, E211Q, E211R, E211S, E211T, E211V,E211W, E211Y, K212*, K212A, K212D, K212E, K212F, K212G, K212H, K212I,K212L, K212M, K212N, K212P, K212Q, K212R, K212S, K212T, K212V, K212W,K212Y, N213*, N213A, N213D, N213E, N213F, N213G, N213H, N213I, N213K,N213L, N213M, N213P, N213Q, N213R, N213S, N213T, N213V, N213W, N213Y,N214*, N214A, N214D, N214E, N214F, N214G, N214H, N214I, N214K, N214L,N214M, N214P, N214Q, N214R, N214S, N214T, N214V, N214W, N214Y, K215*,K215A, K215D, K215E, K215F, K215G, K215H, K215I, K215L, K215M, K215N,K215P, K215Q, K215R, K215S, K215T, K215V, K215W, K215Y, F216*, F216A,F216D, F216E, F216G, F216H, F216I, F216K, F216L, F216M, F216N, F216P,F216Q, F216R, F216S, F216T, F216V, F216W, F216Y, N217*, N217A, N217D,N217E, N217F, N217G, N217H, N217I, N217K, N217L, N217M, N217P, N217Q,N217R, N217S, N217T, N217V, N217W, N217Y, Y218*, Y218A, Y218D, Y218E,Y218F, Y218G, Y218H, Y218I, Y218K, Y218L, Y218M, Y218N, Y218P, Y218Q,Y218R, Y218S, Y218T, Y218V, Y218W, V219*, V219A, V219D, V219E, V219F,V219G, V219H, V219I, V219K, V219L, V219M, V219N, V219P, V219Q, V219R,V219S, V219T, V219W, V219Y, G220*, G220A, G220D, G220E, G220F, G220H,G220I, G220K, G220L, G220M, G220N, G220P, G220Q, G220R, G220S, G220T,G220V, G220W, G220Y, K221*, K221A, K221D, K221E, K221F, K221G, K221H,K221I, K221L, K221M, K221N, K221P, K221Q, K221R, K221S, K221T, K221V,K221W and K221Y, wherein each position corresponds to the position ofthe polypeptide of SEQ ID NO: 1.

Some aspect of the invention relates to a variant of a DNase parent,wherein the has least 60%, such as at least 70%, such as at least 80%such as at least 90%, such as at least 95% but less than 100% sequenceidentity to SEQ ID NO: 1, wherein the variant has DNase activity andwherein the variant comprises one or more of the substitutions selectedfrom the group consisting of: V1A, V1D, V1E, V1F, V1G, V1H, V1I, V1K,V1L, V1M, V1N, V1P, V1Q, V1R, V1S, V1T, V1W, V1Y, P2A, P2D, P2E, P2F,P2G, P2H, P2I, P2K, P2L, P2M, P2N, P2Q, P2R, P2S, P2T, P2V, P2W, P2Y,V3A, V3C, V3D, V3E, V3F, V3G, V3H, V3I, V3K, V3L, V3M, V3N, V3P, V3R,V3S, V3T, V3W, V3Y, N4A, N4D, N4E, N4F, N4G, N4H, N4I, N4K, N4L, N4M,N4P, N4Q, N4R, N4S, N4T, N4V, N4W, N4Y, P5A, P5D, P5E, P5F, P5G, P5H,P5I, P5K, P5L, P5M, P5N, P5Q, P5R, P5S, P5T, P5V, P5W, P5Y, E6A, E6D,E6F, E6G, E6H, E6I, E6K, E6L, E6M, E6N, E6P, E6Q, E6R, E6S, E6T, E6V,E6W, E6Y, P7A, P7D, P7E, P7F, P7G, P7H, P7I, P7K, P7L, P7M, P7N, P7Q,P7R, P7S, P7T, P7V, P7W, P7Y, D8A, D8E, D8F, D8G, D8H, D8I, D8K, D8L,D8M, D8N, D8P, D8Q, D8R, D8S, D8T, D8V, D8W, D8Y, A9D, A9E, A9F, A9G,A9H, A9I, A9K, A9L, A9M, A9N, A9P, A9Q, A9R, A9S, A9T, A9V, A9W, A9Y,T10A, T10D, T10E, T10F, T10G, T10H, T10I, T10K, T10L, T10M, T10N, T10P,T10Q, T10R, T10S, T10V, T10W, T10Y, S11A, S11D, S11E, S11F, S11G, S11H,S11I, S11K, S11L, S11M, S11N, S11P, S11Q, S11R, S11T, S11V, S11W, S11Y,V12A, V12D, V12E, V12F, V12G, V12H, V12I, V12K, V12L, V12M, V12N, V12P,V12Q, V12R, V12S, V12T, V12W, V12Y, E13A, E13D, E13F, E13G, E13H, E13I,E13K, E13L, E13M, E13N, E13P, E13Q, E13R, E13S, E13T, E13V, E13W, E13Y,N14A, N14D, N14E, N14F, N14G, N14H, N14I, N14K, N14L, N14M, N14P, N14Q,N14R, N14S, N14T, N14V, N14W, N14Y, V15A, V15D, V15E, V15F, V15G, V15H,V15I, V15K, V15L, V15M, V15N, V15P, V15Q, V15R, V15S, V15T, V15W, V15Y,A16D, A16E, A16F, A16G, A16H, A16I, A16K, A16L, A16M, A16N, A16P, A16Q,A16R, A16S, A16T, A16V, A16W, A16Y, L17A, L17D, L17E, L17F, L17G, L17H,L17I, L17K, L17M, L17N, L17P, L17Q, L17R, L17S, L17T, L17V, L17W, L17Y,K18A, K18D, K18E, K18F, K18G, K18H, K18I, K18L, K18M, K18N, K18P, K18Q,K18R, K18S, K18T, K18V, K18W, K18Y, T19A, T19D, T19E, T19F, T19G, T19H,T19I, T19K, T19L, T19M, T19N, T19P, T19Q, T19R, T19S, T19V, T19W, T19Y,G20A, G20D, G20E, G20F, G20H, G20I, G20K, G20L, G20M, G20N, G20P, G20Q,G20R, G20S, G20T, G20V, G20W, G20Y, S21A, S21D, S21E, S21F, S21G, S21H,S21I, S21K, S21L, S21M, S21N, S21P, S21Q, S21R, S21T, S21V, S21W, S21Y,G22A, G22D, G22E, G22F, G22H, G22I, G22K, G22L, G22M, G22N, G22P, G22Q,G22R, G22S, G22T, G22V, G22W, G22Y, D23A, D23E, D23F, D23G, D23H, D23I,D23K, D23L, D23M, D23N, D23P, D23Q, D23R, D23S, D23T, D23V, D23W, D23Y,S24A, S24D, S24E, S24F, S24G, S24H, S24I, S24K, S24L, S24M, S24N, S24P,S24Q, S24R, S24T, S24V, S24W, S24Y, Q25A, Q25D, Q25E, Q25F, Q25G, Q25H,Q25I, Q25K, Q25L, Q25M, Q25N, Q25P, Q25R, Q25S, Q25T, Q25V, Q25W, Q25Y,S26A, S26D, S26E, S26F, S26G, S26H, S26I, S26K, S26L, S26M, S26N, S26P,S26Q, S26R, S26T, S26V, S26W, S26Y, D27A, D27E, D27F, D27G, D27H, D27I,D27K, D27L, D27M, D27N, D27P, D27Q, D27R, D27S, D27T, D27V, D27W, D27Y,P28A, P28D, P28E, P28F, P28G, P28H, P28I, P28K, P28L, P28M, P28N, P28Q,P28R, P28S, P28T, P28V, P28W, P28Y, I29A, I29D, I29E, I29F, I29G, I29H,I29K, I29L, I29M, I29N, I29P, I29Q, I29R, I29S, I29T, I29V, I29W, I29Y,K30A, K30D, K30E, K30F, K30G, K30H, K30I, K30L, K30M, K30N, K30P, K30Q,K30R, K30S, K30T, K30V, K30W, K30Y, A31D, A31E, A31F, A31G, A31H, A31I,A31K, A31L, A31M, A31N, A31P, A31Q, A31R, A31S, A31T, A31V, A31W, A31Y,D32A, D32E, D32F, D32G, D32H, D32I, D32K, D32L, D32M, D32N, D32P, D32Q,D32R, D32S, D32T, D32V, D32W, D32Y, L33A, L33D, L33E, L33F, L33G, L33H,L33I, L33K, L33M, L33N, L33P, L33Q, L33R, L33S, L33T, L33V, L33W, L33Y,E34A, E34D, E34F, E34G, E34H, E34I, E34K, E34L, E34M, E34N, E34P, E34Q,E34R, E34S, E34T, E34V, E34W, E34Y, V35A, V35D, V35E, V35F, V35G, V35H,V35I, V35K, V35L, V35M, V35N, V35P, V35Q, V35R, V35S, V35T, V35W, V35Y,K36A, K36D, K36E, K36F, K36G, K36H, K36I, K36L, K36M, K36N, K36P, K36Q,K36R, K36S, K36T, K36V, K36W, K36Y, G37A, G37D, G37E, G37F, G37H, G37I,G37K, G37L, G37M, G37N, G37P, G37Q, G37R, G37S, G37T, G37V, G37W, G37Y,Q38A, Q38D, Q38E, Q38F, Q38G, Q38H, Q38I, Q38K, Q38L, Q38M, Q38N, Q38P,Q38R, Q38S, Q38T, Q38V, Q38W, Q38Y, S39A, S39D, S39E, S39F, S39G, S39H,S39I, S39K, S39L, S39M, S39N, S39P, S39Q, S39R, S39T, S39V, S39W, S39Y,A40D, A40E, A40F, A40G, A40H, A40I, A40K, A40L, A40M, A40N, A40P, A40Q,A40R, A40S, A40T, A40V, A40W, A40Y, L41A, L41D, L41E, L41F, L41G, L41H,L41I, L41K, L41M, L41N, L41P, L41Q, L41R, L41S, L41T, L41V, L41W, L41Y,P42A, P42D, P42E, P42F, P42G, P42H, P42I, P42K, P42L, P42M, P42N, P42Q,P42R, P42S, P42T, P42V, P42W, P42Y, F43A, F43D, F43E, F43G, F43H, F43I,F43K, F43L, F43M, F43N, F43P, F43Q, F43R, F43S, F43T, F43V, F43W, F43Y,D44A, D44E, D44F, D44G, D44H, D44I, D44K, D44L, D44M, D44N, D44P, D44Q,D44R, D44S, D44T, D44V, D44W, D44Y, V45A, V45D, V45E, V45F, V45G, V45H,V45I, V45K, V45L, V45M, V45N, V45P, V45Q, V45R, V45S, V45T, V45W, V45Y,D46A, D46E, D46F, D46G, D46H, D46I, D46K, D46L, D46M, D46N, D46P, D46Q,D46R, D46S, D46T, D46V, D46W, D46Y, C47A, C47D, C47E, C47F, C47G, C47H,C47I, C47K, C47L, C47M, C47N, C47P, C47Q, C47R, C47S, C47T, C47V, C47W,C47Y, W48A, W48D, W48E, W48F, W48G, W48H, W48I, W48K, W48L, W48M, W48N,W48P, W48Q, W48R, W48S, W48T, W48V, W48Y, A49D, A49E, A49F, A49G, A49H,A49I, A49K, A49L, A49M, A49N, A49P, A49Q, A49R, A49S, A49T, A49V, A49W,A49Y, I50A, I50D, I50E, I50F, I50G, I50H, I50K, I50L, I50M, I50N, I50P,I50Q, I50R, I50S, I50T, I50V, I50W, I50Y, L51A, L51D, L51E, L51F, L51G,L51H, L51I, L51K, L51M, L51N, L51P, L51Q, L51R, L51S, L51T, L51V, L51W,L51Y, K53A, K53D, K53E, K53F, K53G, K53H, K53I, K53L, K53M, K53N, K53P,K53Q, K53R, K53S, K53T, K53V, K53W, K53Y, G54A, G54D, G54E, G54F, G54H,G54I, G54K, G54L, G54M, G54N, G54P, G54Q, G54R, G54S, G54T, G54V, G54W,G54Y, A55D, A55E, A55F, A55G, A55H, A55I, A55K, A55L, A55M, A55N, A55P,A55Q, A55R, A55S, A55T, A55V, A55W, A55Y, P56A, P56D, P56E, P56F, P56G,P56H, P56I, P56K, P56L, P56M, P56N, P56Q, P56R, P56S, P56T, P56V, P56W,P56Y, N57A, N57D, N57E, N57F, N57G, N57H, N57I, N57K, N57L, N57M, N57P,N57Q, N57R, N57S, N57T, N57V, N57W, N57Y, V58A, V58D, V58E, V58F, V58G,V58H, V58I, V58K, V58L, V58M, V58N, V58P, V58Q, V58R, V58S, V58T, V58W,V58Y, L59A, L59D, L59E, L59F, L59G, L59H, L59I, L59K, L59M, L59N, L59P,L59Q, L59R, L59S, L59T, L59V, L59W, L59Y, Q60A, Q60D, Q60E, Q60F, Q60G,Q60H, Q60I, Q60K, Q60L, Q60M, Q60N, Q60P, Q60R, Q60S, Q60T, Q60V, Q60W,Q60Y, R61A, R61D, R61E, R61F, R61G, R61H, R61I, R61K, R61L, R61M, R61N,R61P, R61Q, R61S, R61T, R61V, R61W, R61Y, V62A, V62D, V62E, V62F, V62G,V62H, V62I, V62K, V62L, V62M, V62N, V62P, V62Q, V62R, V62S, V62T, V62W,V62Y, N63A, N63D, N63E, N63F, N63G, N63H, N63I, N63K, N63L, N63M, N63P,N63Q, N63R, N63S, N63T, N63V, N63W, N63Y, E64A, E64D, E64F, E64G, E64H,E64I, E64K, E64L, E64M, E64N, E64P, E64Q, E64R, E64S, E64T, E64V, E64W,E64Y, K65A, K65D, K65E, K65F, K65G, K65H, K65I, K65L, K65M, K65N, K65P,K65Q, K65R, K65S, K65T, K65V, K65W, K65Y, T66A, T66D, T66E, T66F, T66G,T66H, T66I, T66K, T66L, T66M, T66N, T66P, T66Q, T66R, T66S, T66V, T66W,T66Y, K67A, K67D, K67E, K67F, K67G, K67H, K67I, K67L, K67M, K67N, K67P,K67Q, K67R, K67S, K67T, K67V, K67W, K67Y, N68A, N68D, N68E, N68F, N68G,N68H, N68I, N68K, N68L, N68M, N68P, N68Q, N68R, N68S, N68T, N68V, N68W,N68Y, S69A, S69D, S69E, S69F, S69G, S69H, S69I, S69K, S69L, S69M, S69N,S69P, S69Q, S69R, S69T, S69V, S69W, S69Y, N70A, N70D, N70E, N70F, N70G,N70H, N70I, N70K, N70L, N70M, N70P, N70Q, N70R, N70S, N70T, N70V, N70W,N70Y, R71A, R71D, R71E, R71F, R71G, R71H, R71I, R71K, R71L, R71M, R71N,R71P, R71Q, R71S, R71T, R71V, R71W, R71Y, D72A, D72E, D72F, D72G, D72H,D72I, D72K, D72L, D72M, D72N, D72P, D72Q, D72R, D72S, D72T, D72V, D72W,D72Y, R73A, R73D, R73E, R73F, R73G, R73H, R73I, R73K, R73L, R73M, R73N,R73P, R73Q, R73S, R73T, R73V, R73W, R73Y, S74A, S74D, S74E, S74F, S74G,S74H, S74I, S74K, S74L, S74M, S74N, S74P, S74Q, S74R, S74T, S74V, S74W,S74Y, G75A, G75D, G75E, G75F, G75H, G75I, G75K, G75L, G75M, G75N, G75P,G75Q, G75R, G75S, G75T, G75V, G75W, G75Y, A76D, A76E, A76F, A76G, A76H,A76I, A76K, A76L, A76M, A76N, A76P, A76Q, A76R, A76S, A76T, A76V, A76W,A76Y, N77A, N77D, N77E, N77F, N77G, N77H, N77I, N77K, N77L, N77M, N77P,N77Q, N77R, N77S, N77T, N77V, N77W, N77Y, K78A, K78D, K78E, K78F, K78G,K78H, K78I, K78L, K78M, K78N, K78P, K78Q, K78R, K78S, K78T, K78V, K78W,K78Y, G79A, G79D, G79E, G79F, G79H, G79I, G79K, G79L, G79M, G79N, G79P,G79Q, G79R, G79S, G79T, G79V, G79W, G79Y, P80A, P80D, P80E, P80F, P80G,P80H, P80I, P80K, P80L, P80M, P80N, P80Q, P80R, P80S, P80T, P80V, P80W,P80Y, F81A, F81D, F81E, F81G, F81H, F81I, F81K, F81L, F81M, F81N, F81P,F81Q, F81R, F81S, F81T, F81V, F81W, F81Y, K82A, K82D, K82E, K82F, K82G,K82H, K82I, K82L, K82M, K82N, K82P, K82Q, K82R, K82S, K82T, K82V, K82W,K82Y, D83A, D83E, D83F, D83G, D83H, D83I, D83K, D83L, D83M, D83N, D83P,D83Q, D83R, D83S, D83T, D83V, D83W, D83Y, P84A, P84D, P84E, P84F, P84G,P84H, P84I, P84K, P84L, P84M, P84N, P84Q, P84R, P84S, P84T, P84V, P84W,P84Y, Q85A, Q85D, Q85E, Q85F, Q85G, Q85H, Q85I, Q85K, Q85L, Q85M, Q85N,Q85P, Q85R, Q85S, Q85T, Q85V, Q85W, Q85Y, K86A, K86D, K86E, K86F, K86G,K86H, K86I, K86L, K86M, K86N, K86P, K86Q, K86R, K86S, K86T, K86V, K86W,K86Y, W87A, W87D, W87E, W87F, W87G, W87H, W87I, W87K, W87L, W87M, W87N,W87P, W87Q, W87R, W87S, W87T, W87V, W87Y, G88A, G88D, G88E, G88F, G88H,G88I, G88K, G88L, G88M, G88N, G88P, G88Q, G88R, G88S, G88T, G88V, G88W,G88Y, I89A, I89D, 189E, I89F, I89G, I89H, I89K, I89L, I89M, I89N, I89P,I89Q, I89R, I89S, I89T, I89V, I89W, I89Y, K90A, K90D, K90E, K90F, K90G,K90H, K90I, K90L, K90M, K90N, K90P, K90Q, K90R, K90S, K90T, K90V, K90W,K90Y, A91D, A91E, A91F, A91G, A91H, A91I, A91K, A91L, A91M, A91N, A91P,A91Q, A91R, A91S, A91T, A91V, A91W, A91Y, L92A, L92D, L92E, L92F, L92G,L92H, L92I, L92K, L92M, L92N, L92P, L92Q, L92R, L92S, L92T, L92V, L92W,L92Y, P93A, P93D, P93E, P93F, P93G, P93H, P93I, P93K, P93L, P93M, P93N,P93Q, P93R, P93S, P93T, P93V, P93W, P93Y, P94A, P94D, P94E, P94F, P94G,P94H, P94I, P94K, P94L, P94M, P94N, P94Q, P94R, P94S, P94T, P94V, P94W,P94Y, K95A, K95D, K95E, K95F, K95G, K95H, K95I, K95L, K95M, K95N, K95P,K95Q, K95R, K95S, K95T, K95V, K95W, K95Y, N96A, N96D, N96E, N96F, N96G,N96H, N96I, N96K, N96L, N96M, N96P, N96Q, N96R, N96S, N96T, N96V, N96W,N96Y, P97A, P97D, P97E, P97F, P97G, P97H, P97I, P97K, P97L, P97M, P97N,P97Q, P97R, P97S, P97T, P97V, P97W, P97Y, S98A, S98D, S98E, S98F, S98G,S98H, S98I, S98K, S98L, S98M, S98N, S98P, S98Q, S98R, S98T, S98V, S98W,S98Y, W99A, W99D, W99E, W99F, W99G, W99H, W99I, W99K, W99L, W99M, W99N,W99P, W99Q, W99R, W99S, W99T, W99V, W99Y, S100A, S100D, S100E, S100F,S100G, S100H, S100I, S100K, S100L, S100M, S100N, S100P, S100Q, S100R,S100T, S100V, S100W, S100Y, A101D, A101E, A101F, A101G, A101H, A101I,A101K, A101L, A101M, A101N, A101P, A101Q, A101R, A101S, A101T, A101V,A101W, A101Y, Q102A, Q102D, Q102E, Q102F, Q102G, Q102H, Q102I, Q102K,Q102L, Q102M, Q102N, Q102P, Q102R, Q102S, Q102T, Q102V, Q102W, Q102Y,D103A, D103E, D103F, D103G, D103H, D103I, D103K, D103L, D103M, D103N,D103P, D103Q, D103R, D103S, D103T, D103V, D103W, D103Y, F104A, F104D,F104E, F104G, F104H, F104I, F104K, F104L, F104M, F104N, F104P, F104Q,F104R, F104S, F104T, F104V, F104W, F104Y, K105A, K105D, K105E, K105F,K105G, K105H, K105I, K105L, K105M, K105N, K105P, K105Q, K105R, K105S,K105T, K105V, K105W, K105Y, S106A, S106D, S106E, S106F, S106G, S106H,S106I, S106K, S106L, S106M, S106N, S106P, S106Q, S106R, S106T, S106V,S106W, S106Y, P107A, P107D, P107E, P107F, P107G, P107H, P107I, P107K,P107L, P107M, P107N, P107Q, P107R, P107S, P107T, P107V, P107W, P107Y,E108A, E108D, E108F, E108G, E108H, E108I, E108K, E108L, E108M, E108N,E108P, E108Q, E108R, E108S, E108T, E108V, E108W, E108Y, E109A, E109D,E109F, E109G, E109H, E109I, E109K, E109L, E109M, E109N, E109P, E109Q,E109R, E109S, E109T, E109V, E109W, E109Y, Y110A, Y110D, Y110E, Y110F,Y110G, Y110H, Y110I, Y110K, Y110L, Y110M, Y110N, Y110P, Y110Q, Y110R,Y110S, Y110T, Y110V, Y110W, A111D, A111E, A111F, A111G, A111H, A111I,A111K, A111L, A111M, A111N, A111P, A111Q, A111R, A111S, A111T, A111V,A111W, A111Y, F112A, F112D, F112E, F112G, F112H, F112I, F112K, F112L,F112M, F112N, F112P, F112Q, F112R, F112S, F112T, F112V, F112W, F112Y,A113D, A113E, A113F, A113G, A113H, A113I, A113K, A113L, A113M, A113N,A113P, A113Q, A113R, A113S, A113T, A113V, A113W, A113Y, S114A, S114D,S114E, S114F, S114G, S114H, S114I, S114K, S114L, S114M, S114N, S114P,S114Q, S114R, S114T, S114V, S114W, S114Y, S115A, S115D, S115E, S115F,S115G, S115H, S115I, S115K, S115L, S115M, S115N, S115P, S115Q, S115R,S115T, S115V, S115W, S115Y, L116A, L116D, L116E, L116F, L116G, L116H,L116I, L116K, L116M, L116N, L116P, L116Q, L116R, L116S, L116T, L116V,L116W, L116Y, Q117A, Q117D, Q117E, Q117F, Q117G, Q117H, Q117I, Q117K,Q117L, Q117M, Q117N, Q117P, Q117R, Q117S, Q117T, Q117V, Q117W, Q117Y,G118A, G118D, G118E, G118F, G118H, G118I, G118K, G118L, G118M, G118N,G118P, G118Q, G118R, G118S, G118T, G118V, G118W, G118Y, G119A, G119D,G119E, G119F, G119H, G119I, G119K, G119L, G119M, G119N, G119P, G119Q,G119R, G119S, G119T, G119V, G119W, G119Y, T120A, T120D, T120E, T120F,T120G, T120H, T120I, T120K, T120L, T120M, T120N, T120P, T120Q, T120R,T120S, T120V, T120W, T120Y, N121A, N121D, N121E, N121F, N121G, N121H,N121I, N121K, N121L, N121M, N121P, N121Q, N121R, N121S, N121T, N121V,N121W, N121Y, A122D, A122E, A122F, A122G, A122H, A122I, A122K, A122L,A122M, A122N, A122P, A122Q, A122R, A122S, A122T, A122V, A122W, A122Y,I123A, I123D, I123E, I123F, I123G, I123H, I123K, I123L, I123M, I123N,I123P, I123Q, I123R, I123S, I123T, I123V, I123W, I123Y, L124A, L124D,L124E, L124F, L124G, L124H, L124I, L124K, L124M, L124N, L124P, L124Q,L124R, L124S, L124T, L124V, L124W, L124Y, A125D, A125E, A125F, A125G,A125H, A125I, A125K, A125L, A125M, A125N, A125P, A125Q, A125R, A125S,A125T, A125V, A125W, A125Y, P126A, P126D, P126E, P126F, P126G, P126H,P126I, P126K, P126L, P126M, P126N, P126Q, P126R, P126S, P126T, P126V,P126W, P126Y, V127A, V127D, V127E, V127F, V127G, V127H, V127I, V127K,V127L, V127M, V127N, V127P, V127Q, V127R, V127S, V127T, V127W, V127Y,N128A, N128D, N128E, N128F, N128G, N128H, N128I, N128K, N128L, N128M,N128P, N128Q, N128R, N128S, N128T, N128V, N128W, N128Y, L129A, L129D,L129E, L129F, L129G, L129H, L129I, L129K, L129M, L129N, L129P, L129Q,L129R, L129S, L129T, L129V, L129W, L129Y, A130D, A130E, A130F, A130G,A130H, A130I, A130K, A130L, A130M, A130N, A130P, A130Q, A130R, A130S,A130T, A130V, A130W, A130Y, S131A, S131D, S131E, S131F, S131G, S131H,S131I, S131K, S131L, S131M, S131N, S131P, S131Q, S131R, S131T, S131V,S131W, S131Y, Q132A, Q132D, Q132E, Q132F, Q132G, Q132H, Q132I, Q132K,Q132L, Q132M, Q132N, Q132P, Q132R, Q132S, Q132T, Q132V, Q132W, Q132Y,N133A, N133D, N133E, N133F, N133G, N133H, N133I, N133K, N133L, N133M,N133P, N133Q, N133R, N133S, N133T, N133V, N133W, N133Y, S134A, S134D,S134E, S134F, S134G, S134H, S134I, S134K, S134L, S134M, S134N, S134P,S134Q, S134R, S134T, S134V, S134W, S134Y, Q135A, Q135D, Q135E, Q135F,Q135G, Q135H, Q135I, Q135K, Q135L, Q135M, Q135N, Q135P, Q135R, Q135S,Q135T, Q135V, Q135W, Q135Y, G136A, G136D, G136E, G136F, G136H, G136I,G136K, G136L, G136M, G136N, G136P, G136Q, G136R, G136S, G136T, G136V,G136W, G136Y, G137A, G137D, G137E, G137F, G137H, G137I, G137K, G137L,G137M, G137N, G137P, G137Q, G137R, G137S, G137T, G137V, G137W, G137Y,V138A, V138D, V138E, V138F, V138G, V138H, V138I, V138K, V138L, V138M,V138N, V138P, V138Q, V138R, V138S, V138T, V138W, V138Y, L139A, L139D,L139E, L139F, L139G, L139H, L139I, L139K, L139M, L139N, L139P, L139Q,L139R, L139S, L139T, L139V, L139W, L139Y, N140A, N140D, N140E, N140F,N140G, N140H, N140I, N140K, N140L, N140M, N140P, N140Q, N140R, N140S,N140T, N140V, N140W, N140Y, G141A, G141D, G141E, G141F, G141H, G141I,G141K, G141L, G141M, G141N, G141P, G141Q, G141R, G141S, G141T, G141V,G141W, G141Y, F142A, F142D, F142E, F142G, F142H, F142I, F142K, F142L,F142M, F142N, F142P, F142Q, F142R, F142S, F142T, F142V, F142W, F142Y,Y143A, Y143D, Y143E, Y143F, Y143G, Y143H, Y143I, Y143K, Y143L, Y143M,Y143N, Y143P, Y143Q, Y143R, Y143S, Y143T, Y143V, Y143W, S144A, S144D,S144E, S144F, S144G, S144H, S144I, S144K, S144L, S144M, S144N, S144P,S144Q, S144R, S144T, S144V, S144W, S144Y, A145D, A145E, A145F, A145G,A145H, A145I, A145K, A145L, A145M, A145N, A145P, A145Q, A145R, A145S,A145T, A145V, A145W, A145Y, N146A, N146D, N146E, N146F, N146G, N146H,N146I, N146K, N146L, N146M, N146P, N146Q, N146R, N146S, N146T, N146V,N146W, N146Y, K147A, K147D, K147E, K147F, K147G, K147H, K147I, K147L,K147M, K147N, K147P, K147Q, K147R, K147S, K147T, K147V, K147W, K147Y,V148A, V148D, V148E, V148F, V148G, V148H, V148I, V148K, V148L, V148M,V148N, V148P, V148Q, V148R, V148S, V148T, V148W, V148Y, A149D, A149E,A149F, A149G, A149H, A149I, A149K, A149L, A149M, A149N, A149P, A149Q,A149R, A149S, A149T, A149V, A149W, A149Y, Q150A, Q150D, Q150E, Q150F,Q150G, Q150H, Q150I, Q150K, Q150L, Q150M, Q150N, Q150P, Q150R, Q150S,Q150T, Q150V, Q150W, Q150Y, F151A, F151D, F151E, F151G, F151H, F151I,F151K, F151L, F151M, F151N, F151P, F151Q, F151R, F151S, F151T, F151V,F151W, F151Y, D152A, D152E, D152F, D152G, D152H, D152I, D152K, D152L,D152M, D152N, D152P, D152Q, D152R, D152S, D152T, D152V, D152W, D152Y,P153A, P153D, P153E, P153F, P153G, P153H, P153I, P153K, P153L, P153M,P153N, P153Q, P153R, P153S, P153T, P153V, P153W, P153Y, S154A, S154D,S154E, S154F, S154G, S154H, S154I, S154K, S154L, S154M, S154N, S154P,S154Q, S154R, S154T, S154V, S154W, S154Y, K155A, K155D, K155E, K155F,K155G, K155H, K155I, K155L, K155M, K155N, K155P, K155Q, K155R, K155S,K155T, K155V, K155W, K155Y, P156A, P156D, P156E, P156F, P156G, P156H,P156I, P156K, P156L, P156M, P156N, P156Q, P156R, P156S, P156T, P156V,P156W, P156Y, Q157A, Q157D, Q157E, Q157F, Q157G, Q157H, Q157I, Q157K,Q157L, Q157M, Q157N, Q157P, Q157R, Q157S, Q157T, Q157V, Q157W, Q157Y,Q158A, Q158D, Q158E, Q158F, Q158G, Q158H, Q158I, Q158K, Q158L, Q158M,Q158N, Q158P, Q158R, Q158S, Q158T, Q158V, Q158W, Q158Y, T159A, T159D,T159E, T159F, T159G, T159H, T159I, T159K, T159L, T159M, T159N, T159P,T159Q, T159R, T159S, T159V, T159W, T159Y, K160A, K160D, K160E, K160F,K160G, K160H, K160I, K160L, K160M, K160N, K160P, K160Q, K160R, K160S,K160T, K160V, K160W, K160Y, G161A, G161D, G161E, G161F, G161H, G161I,G161K, G161L, G161M, G161N, G161P, G161Q, G161R, G161S, G161T, G161V,G161W, G161Y, T162A, T162D, T162E, T162F, T162G, T162H, T162I, T162K,T162L, T162M, T162N, T162P, T162Q, T162R, T162S, T162T, T162V, T162W,T162Y, W163A, W163D, W163E, W163F, W163G, W163H, W163I, W163K, W163L,W163M, W163N, W163P, W163Q, W163R, W163S, W163T, W163V, W163Y, F164A,F164D, F164E, F164G, F164H, F164I, F164K, F164L, F164M, F164N, F164P,F164Q, F164R, F164S, F164T, F164V, F164W, F164Y, Q165A, Q165D, Q165E,Q165F, Q165G, Q165H, Q165I, Q165K, Q165L, Q165M, Q165N, Q165P, Q165R,Q165S, Q165T, Q165V, Q165W, Q165Y, I166A, I166D, I166E, I166F, I166G,I166H, I166K, I166L, I166M, I166N, I166P, I166Q, I166R, I166S, I166T,I166V, I166W, I166Y, T167A, T167D, T167E, T167F, T167G, T167H, T167I,T167K, T167L, T167M, T167N, T167P, T167Q, T167R, T167S, T167V, T167W,T167Y, K168A, K168D, K168E, K168F, K168G, K168H, K168I, K168L, K168M,K168N, K168P, K168Q, K168R, K168S, K168T, K168V, K168W, K168Y, F169A,F169D, F169E, F169G, F169H, F169I, F169K, F169L, F169M, F169N, F169P,F169Q, F169R, F169S, F169T, F169V, F169W, F169Y, T170A, T170D, T170E,T170F, T170G, T170H, T170I, T170K, T170L, T170M, T170N, T170P, T170Q,T170R, T170S, T170V, T170W, T170Y, G171A, G171D, G171E, G171F, G171H,G171I, G171K, G171L, G171M, G171N, G171P, G171Q, G171R, G171S, G171T,G171V, G171W, G171Y, A172D, A172E, A172F, A172G, A172H, A172I, A172K,A172L, A172M, A172N, A172P, A172Q, A172R, A172S, A172T, A172V, A172W,A172Y, A173D, A173E, A173F, A173G, A173H, A173I, A173K, A173L, A173M,A173N, A173P, A173Q, A173R, A173S, A173T, A173V, A173W, A173Y, G174A,G174D, G174E, G174F, G174H, G174I, G174K, G174L, G174M, G174N, G174P,G174Q, G174R, G174S, G174T, G174V, G174W, G174Y, P175A, P175D, P175E,P175F, P175G, P175H, P175I, P175K, P175L, P175M, P175N, P175Q, P175R,P175S, P175T, P175V, P175W, P175Y, Y176A, Y176D, Y176E, Y176F, Y176G,Y176H, Y176I, Y176K, Y176L, Y176M, Y176N, Y176P, Y176Q, Y176R, Y176S,Y176T, Y176V, Y176W, K178A, K178D, K178E, K178F, K178G, K178H, K178I,K178L, K178M, K178N, K178P, K178Q, K178R, K178S, K178T, K178V, K178W,K178Y, A179D, A179E, A179F, A179G, A179H, A179I, A179K, A179L, A179M,A179N, A179P, A179Q, A179R, A179S, A179T, A179V, A179W, A179Y, L180A,L180D, L180E, L180F, L180G, L180H, L180I, L180K, L180M, L180N, L180P,L180Q, L180R, L180S, L180T, L180V, L180W, L180Y, G181A, G181D, G181E,G181F, G181H, G181I, G181K, G181L, G181M, G181N, G181P, G181Q, G181R,G181S, G181T, G181V, G181W, G181Y, S182A, S182D, S182E, S182F, S182G,S182H, S182I, S182K, S182L, S182M, S182N, S182P, S182Q, S182R, S182T,S182V, S182W, S182Y, N183A, N183D, N183E, N183F, N183G, N183H, N183I,N183K, N183L, N183M, N183P, N183Q, N183R, N183S, N183T, N183V, N183W,N183Y, D184A, D184E, D184F, D184G, D184H, D184I, D184K, D184L, D184M,D184N, D184P, D184Q, D184R, D184S, D184T, D184V, D184W, D184Y, K185A,K185D, K185E, K185F, K185G, K185H, K185I, K185L, K185M, K185N, K185P,K185Q, K185R, K185S, K185T, K185V, K185W, K185Y, S186A, S186D, S186E,S186F, S186G, S186H, S186I, S186K, S186L, S186M, S186N, S186P, S186Q,S186R, S186T, S186V, S186W, S186Y, V187A, V187D, V187E, V187F, V187G,V187H, V187I, V187K, V187L, V187M, V187N, V187P, V187Q, V187R, V187S,V187T, V187W, V187Y, D189A, D189E, D189F, D189G, D189H, D189I, D189K,D189L, D189M, D189N, D189P, D189Q, D189R, D189S, D189T, D189V, D189W,D189Y, K190A, K190D, K190E, K190F, K190G, K190H, K190I, K190L, K190M,K190N, K190P, K190Q, K190R, K190S, K190T, K190V, K190W, K190Y, N191A,N191D, N191E, N191F, N191G, N191H, N191I, N191K, N191L, N191M, N191P,N191Q, N191R, N191S, N191T, N191V, N191W, N191Y, K192A, K192D, K192E,K192F, K192G, K192H, K192I, K192L, K192M, K192N, K192P, K192Q, K192R,K192S, K192T, K192V, K192W, K192Y, N193A, N193D, N193E, N193F, N193G,N193H, N193I, N193K, N193L, N193M, N193P, N193Q, N193R, N193S, N193T,N193V, N193W, N193Y, I194A, I194D, I194E, I194F, I194G, I194H, I194K,I194L, I194M, I194N, I194P, I194Q, I194R, I194S, I194T, I194V, I194W,I194Y, A195D, A195E, A195F, A195G, A195H, A195I, A195K, A195L, A195M,A195N, A195P, A195Q, A195R, A195S, A195T, A195V, A195W, A195Y, G196A,G196D, G196E, G196F, G196H, G196I, G196K, G196L, G196M, G196N, G196P,G196Q, G196R, G196S, G196T, G196V, G196W, G196Y, D197A, D197E, D197F,D197G, D197H, D197I, D197K, D197L, D197M, D197N, D197P, D197Q, D197R,D197S, D197T, D197V, D197W, D197Y, W198A, W198D, W198E, W198F, W198G,W198H, W198I, W198K, W198L, W198M, W198N, W198P, W198Q, W198R, W198S,W198T, W198V, W198Y, G199A, G199D, G199E, G199F, G199H, G199I, G199K,G199L, G199M, G199N, G199P, G199Q, G199R, G199S, G199T, G199V, G199W,G199Y, F200A, F200D, F200E, F200G, F200H, F200I, F200K, F200L, F200M,F200N, F200P, F200Q, F200R, F200S, F200T, F200V, F200W, F200Y, D201A,D201E, D201F, D201G, D201H, D201I, D201K, D201L, D201M, D201N, D201P,D201Q, D201R, D201S, D201T, D201V, D201W, D201Y, P202A, P202D, P202E,P202F, P202G, P202H, P202I, P202K, P202L, P202M, P202N, P202Q, P202R,P202S, P202T, P202V, P202W, P202Y, A203D, A203E, A203F, A203G, A203H,A203I, A203K, A203L, A203M, A203N, A203P, A203Q, A203R, A203S, A203T,A203V, A203W, A203Y, K204A, K204D, K204E, K204F, K204G, K204H, K204I,K204L, K204M, K204N, K204P, K204Q, K204R, K204S, K204T, K204V, K204W,K204Y, W205A, W205D, W205E, W205F, W205G, W205H, W205I, W205K, W205L,W205M, W205N, W205P, W205Q, W205R, W205S, W205T, W205V, W205Y, A206D,A206E, A206F, A206G, A206H, A206I, A206K, A206L, A206M, A206N, A206P,A206Q, A206R, A206S, A206T, A206V, A206W, A206Y, Y207A, Y207D, Y207E,Y207F, Y207G, Y207H, Y207I, Y207K, Y207L, Y207M, Y207N, Y207P, Y207Q,Y207R, Y207S, Y207T, Y207V, Y207W, Q208A, Q208D, Q208E, Q208F, Q208G,Q208H, Q208I, Q208K, Q208L, Q208M, Q208N, Q208P, Q208R, Q208S, Q208T,Q208V, Q208W, Q208Y, Y209A, Y209D, Y209E, Y209F, Y209G, Y209H, Y209I,Y209K, Y209L, Y209M, Y209N, Y209P, Y209Q, Y209R, Y209S, Y209T, Y209V,Y209W, D210A, D210E, D210F, D210G, D210H, D210I, D210K, D210L, D210M,D210N, D210P, D210Q, D210R, D210S, D210T, D210V, D210W, D210Y, E211A,E211D, E211F, E211G, E211H, E211I, E211K, E211L, E211M, E211N, E211P,E211Q, E211R, E211S, E211T, E211V, E211W, E211Y, K212A, K212D, K212E,K212F, K212G, K212H, K212I, K212L, K212M, K212N, K212P, K212Q, K212R,K212S, K212T, K212V, K212W, K212Y, N213A, N213D, N213E, N213F, N213G,N213H, N213I, N213K, N213L, N213M, N213P, N213Q, N213R, N213S, N213T,N213V, N213W, N213Y, N214A, N214D, N214E, N214F, N214G, N214H, N214I,N214K, N214L, N214M, N214P, N214Q, N214R, N214S, N214T, N214V, N214W,N214Y, K215A, K215D, K215E, K215F, K215G, K215H, K215I, K215L, K215M,K215N, K215P, K215Q, K215R, K215S, K215T, K215V, K215W, K215Y, F216A,F216D, F216E, F216G, F216H, F216I, F216K, F216L, F216M, F216N, F216P,F216Q, F216R, F216S, F216T, F216V, F216W, F216Y, N217A, N217D, N217E,N217F, N217G, N217H, N217I, N217K, N217L, N217M, N217P, N217Q, N217R,N217S, N217T, N217V, N217W, N217Y, Y218A, Y218D, Y218E, Y218F, Y218G,Y218H, Y218I, Y218K, Y218L, Y218M, Y218N, Y218P, Y218Q, Y218R, Y218S,Y218T, Y218V, Y218W, V219A, V219D, V219E, V219F, V219G, V219H, V219I,V219K, V219L, V219M, V219N, V219P, V219Q, V219R, V219S, V219T, V219W,V219Y, G220A, G220D, G220E, G220F, G220H, G220I, G220K, G220L, G220M,G220N, G220P, G220Q, G220R, G220S, G220T, G220V, G220W, G220Y, K221A,K221D, K221E, K221F, K221G, K221H, K221I, K221L, K221M, K221N, K221P,K221Q, K221R, K221S, K221T, K221V, K221W and K221Y, wherein eachposition corresponds to the position of the polypeptide of SEQ ID NO: 1.

Some preferred aspect of the invention relates to a DNase variant havingDNase activity, wherein the variant comprise one or more substitutionsselected from the group consisting of: N4E, L17E, T19A, T19G, T19I,K36P, Q38P, S39V, S39R, A40P, A40H, L41T, L41H, V45H, L51G, K53T, K53P,G54P, A55P, N57H, E64A, E64Q, E64R, E64T, E64I, E64S, T66H, K67A, K67T,N68V, N68P, N68I, N68H, S69A, S69D, S69E, S69K, S69L, S69W, S69Y, S69Q,N70T, N70H, N70G, R71T, D72E, S74H, S74G, G75I, N77T, K82P, K82I, D83T,D83P, D83I, D83H, D83G, P84H, Q85T, Q85P, Q85H, K86T, K86P, K86H, G88P,G88H, A91P, W99T, A101W, K105E, K105N, K105T, K105D, S106T, S115T,L116I, Q135L, G136L, V138I, V138L, V138P, V138Q, L139A, N140R, N140L,N140A, G141L, F151R, D152Y, D152L, D152I, D152A, P153E, S154R, T162R,W163E, F164R, I166Y, I166R, K168N, F169R, F169E, A173I, A173R, A173T,S182R, N183E, D184I, K185Y, S186I, D189G, D189H, K212G, K212P and K215I,wherein each position corresponds to the position of the polypeptide ofSEQ ID NO: 1 and wherein the variant has at least 80% sequence identityto the polypeptide shown in SEQ ID NO 1.

In one preferred aspect the invention relates to a DNase variantcomprising one or more deletion compared to SEQ ID NO 1, wherein theamino acid in any of the position selected from positions 84, 88, 139,179 and 180 is deleted.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 1 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 1 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Thr, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from V1A, V1D, V1E, V1F,V1G, V1H, V1I, V1K, V1L, V1M, V1N, V1P, V1Q, V1R, V1S, V1T, V1W and V1Yof the polypeptide shown in SEQ ID NO: 1 or a polypeptide having atleast 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 2 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 2 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg,Ser, Val, Thr, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from P2A, P2D, P2E, P2F,P2G, P2H, P2I, P2K, P2L, P2M, P2N, P2Q, P2R, P2S, P2T, P2V, P2W and P2Yof the polypeptide shown in SEQ ID NO: 1 or a polypeptide having atleast 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 3 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 3 is substitutedwith Ala, Cys, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Arg, Ser, Thr, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from V3A, V3C, V3D, V3E,V3F, V3G, V3H, V3I, V3K, V3L, V3M, V3N, V3P, V3R, V3S, V3T, V3W and V3Yof the polypeptide shown in SEQ ID NO: 1 or a polypeptide having atleast 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 4 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 4 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from N4A, N4D, N4E, N4F,N4G, N4H, N4I, N4K, N4L, N4M, N4P, N4Q, N4R, N4S, N4T, N4V, N4W and N4Yof the polypeptide shown in SEQ ID NO: 1 or a polypeptide having atleast 80% sequence identity hereto. In a preferred aspect, the variantcomprises or consists of the substitution N4E of the polypeptide shownin SEQ ID NO: 1 or a polypeptide having at least 80% sequence identityhereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 5 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 5 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from P5A, P5D, P5E, P5F,P5G, P5H, P5I, P5K, P5L, P5M, P5N, P5Q, P5R, P5S, P5T, P5V, P5W and P5Yof the polypeptide shown in SEQ ID NO: 1 or a polypeptide having atleast 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 6 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 6 is substitutedwith Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from E6A, E6D, E6F, E6G,E6H, E6I, E6K, E6L, E6M, E6N, E6P, E6Q, E6R, E6S, E6T, E6V, E6W, E6Y ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 7 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 7 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from P7A, P7D, P7E, P7F,P7G, P7H, P7I, P7K, P7L, P7M, P7N, P7Q, P7R, P7S, P7T, P7V, P7W and P7Yof the polypeptide shown in SEQ ID NO: 1 or a polypeptide having atleast 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 8 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 8 is substitutedwith Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from D8A, D8E, D8F, D8G,D8H, D8I, D8K, D8L, D8M, D8N, D8P, D8Q, D8R, D8S, D8T, D8V, D8W and D8Yof the polypeptide shown in SEQ ID NO: 1 or a polypeptide having atleast 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 9 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 9 is substitutedwith Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from A9D, A9E, A9F, A9G,A9H, A9I, A9K, A9L, A9M, A9N, A9P, A9Q, A9R, A9S, A9T, A9V, A9W and A9Yof the polypeptide shown in SEQ ID NO: 1 or a polypeptide having atleast 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 10 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 10 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from T10A, T10D, T10E,T10F, T10G, T10H, T10I, T10K, T10L, T10M, T10N, T10P, T10Q, T10R, T10S,T10V, T10W and T10Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 11 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 11 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from S11A, S11D, S11E,S11F, S11G, S11H, S11I, S11K, S11L, S11M, S11N, S11P, S11Q, S11R, S11T,S11V, S11W and S11Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 12 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 12 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Thr, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from V12A, V12D, V12E,V12F, V12G, V12H, V12I, V12K, V12L, V12M, V12N, V12P, V12Q, V12R, V12S,V12T, V12W and V12Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 13 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 13 is substitutedwith Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from E13A, E13D, E13F,E13G, E13H, E13I, E13K, E13L, E13M, E13N, E13P, E13Q, E13R, E13S, E13T,E13V, E13W and E13Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 14 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 14 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from N14A, N14D, N14E,N14F, N14G, N14H, N14I, N14K, N14L, N14M, N14P, N14Q, N14R, N14S, N14T,N14V, N14W and N14Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 15 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 15 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Thr, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from V15A, V15D, V15E,V15F, V15G, V15H, V15I, V15K, V15L, V15M, V15N, V15P, V15Q, V15R, V15S,V15T, V15W and V15Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 16 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 16 is substitutedwith Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from A16D, A16E, A16F,A16G, A16H, A16I, A16K, A16L, A16M, A16N, A16P, A16Q, A16R, A16S, A16T,A16V, A16W and A16Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 17 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 17 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from L17A, L17D, L17E,L17F, L17G, L17H, L17I, L17K, L17M, L17N, L17P, L17Q, L17R, L17S, L17T,L17V, L17W and L17Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution L17E ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 18 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 18 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from K18A, K18D, K18E,K18F, K18G, K18H, K18I, K18L, K18M, K18N, K18P, K18Q, K18R, K18S, K18T,K18V, K18W and K18Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 19 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 19 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from T19A, T19D, T19E,T19F, T19G, T19H, T19I, T19K, T19L, T19M, T19N, T19P, T19Q, T19R, T19S,T19V, T19W and T19Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsT19A, T19G or T19I of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 20 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 20 is substitutedwith Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from G20A, G20D, G20E,G20F, G20H, G20I, G20K, G20L, G20M, G20N, G20P, G20Q, G20R, G20S, G20T,G20V, G20W and G20Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 21 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 21 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from S21A, S21D, S21E,S21F, S21G, S21H, S21I, S21K, S21L, S21M, S21N, S21P, S21Q, S21R, S21T,S21V, S21W and S21Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 22 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 22 is substitutedwith Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from G22A, G22D, G22E,G22F, G22H, G22I, G22K, G22L, G22M, G22N, G22P, G22Q, G22R, G22S, G22T,G22V, G22W and G22Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 23 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 23 is substitutedwith Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from D23A, D23E, D23F,D23G, D23H, D23I, D23K, D23L, D23M, D23N, D23P, D23Q, D23R, D23S, D23T,D23V, D23W and D23Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 24 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 24 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from S24A, S24D, S24E,S24F, S24G, S24H, S24I, S24K, S24L, S24M, S24N, S24P, S24Q, S24R, S24T,S24V, S24W and S24Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 25 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 25 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from Q25A, Q25D, Q25E,Q25F, Q25G, Q25H, Q25I, Q25K, Q25L, Q25M, Q25N, Q25P, Q25R, Q25S, Q25T,Q25V, Q25W and Q25Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 26 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 26 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from S26A, S26D, S26E,S26F, S26G, S26H, S26I, S26K, S26L, S26M, S26N, S26P, S26Q, S26R, S26T,S26V, S26W and S26Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 27 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 27 is substitutedwith Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from D27A, D27E, D27F,D27G, D27H, D27I, D27K, D27L, D27M, D27N, D27P, D27Q, D27R, D27S, D27T,D27V, D27W and D27Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 28 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 28 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from P28A, P28D, P28E,P28F, P28G, P28H, P28I, P28K, P28L, P28M, P28N, P28Q, P28R, P28S, P28T,P28V, P28W and P28Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 29 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 29 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from 129A, I29D, I29E,I29F, I29G, I29H, I29K, I29L, I29M, I29N, I29P, I29Q, I29R, I29S, I29T,I29V, I29W and 129Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 30 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 30 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from K30A, K30D, K30E,K30F, K30G, K30H, K30I, K30L, K30M, K30N, K30P, K30Q, K30R, K30S, K30T,K30V, K30W and K30Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 31 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 31 is substitutedwith Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from A31D, A31E, A31F,A31G, A31H, A31I, A31K, A31L, A31M, A31N, A31P, A31Q, A31R, A31S, A31T,A31V, A31W and A31Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 32 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 32 is substitutedwith Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from D32A, D32E, D32F,D32G, D32H, D32I, D32K, D32L, D32M, D32N, D32P, D32Q, D32R, D32S, D32T,D32V, D32W and D32Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 33 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 33 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from L33A, L33D, L33E,L33F, L33G, L33H, L33I, L33K, L33M, L33N, L33P, L33Q, L33R, L33S, L33T,L33V, L33W and L33Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 34 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 34 is substitutedwith Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from E34A, E34D, E34F,E34G, E34H, E34I, E34K, E34L, E34M, E34N, E34P, E34Q, E34R, E34S, E34T,E34V, E34W and E34Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 35 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 35 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Thr, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from V35A, V35D, V35E,V35F, V35G, V35H, V35I, V35K, V35L, V35M, V35N, V35P, V35Q, V35R, V35S,V35T, V35W and V35Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 36 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 36 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from K36A, K36D, K36E,K36F, K36G, K36H, K36I, K36L, K36M, K36N, K36P, K36Q, K36R, K36S, K36T,K36V, K36W and K36Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution K36P ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 37 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 37 is substitutedwith Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from G37A, G37D, G37E,G37F, G37H, G37I, G37K, G37L, G37M, G37N, G37P, G37Q, G37R, G37S, G37T,G37V, G37W and G37Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 38 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 38 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from Q38A, Q38D, Q38E,Q38F, Q38G, Q38H, Q38I, Q38K, Q38L, Q38M, Q38N, Q38P, Q38R, Q38S, Q38T,Q38V, Q38W and Q38Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution Q38P ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 39 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 39 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from S39A, S39D, S39E,S39F, S39G, S39H, S39I, S39K, S39L, S39M, S39N, S39P, S39Q, S39R, S39T,S39V, S39W and S39Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsS39V or S39R of the polypeptide shown in SEQ ID NO: 1 or a polypeptidehaving at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 40 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 40 is substitutedwith Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from A40D, A40E, A40F,A40G, A40H, A40I, A40K, A40L, A40M, A40N, A40P, A40Q, A40R, A40S, A40T,A40V, A40W, A40Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsA40P or A40H of the polypeptide shown in SEQ ID NO: 1 or a polypeptidehaving at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 41 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 41 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from L41A, L41D, L41E,L41F, L41G, L41H, L41I, L41K, L41M, L41N, L41P, L41Q, L41R, L41S, L41T,L41V, L41W and L41Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsL41T or L41H of the polypeptide shown in SEQ ID NO: 1 or a polypeptidehaving at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 42 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 42 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from P42A, P42D, P42E,P42F, P42G, P42H, P42I, P42K, P42L, P42M, P42N, P42Q, P42R, P42S, P42T,P42V, P42W and P42Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 43 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 43 is substitutedwith Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from F43A, F43D, F43E,F43G, F43H, F43I, F43K, F43L, F43M, F43N, F43P, F43Q, F43R, F43S, F43T,F43V, F43W and F43Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 44 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 44 is substitutedwith Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from D44A, D44E, D44F,D44G, D44H, D44I, D44K, D44L, D44M, D44N, D44P, D44Q, D44R, D44S, D44T,D44V, D44W and D44Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 45 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 45 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Thr, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from V45A, V45D, V45E,V45F, V45G, V45H, V45I, V45K, V45L, V45M, V45N, V45P, V45Q, V45R, V45S,V45T, V45W and V45Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution V45H ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 46 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 46 is substitutedwith Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from D46A, D46E, D46F,D46G, D46H, D46I, D46K, D46L, D46M, D46N, D46P, D46Q, D46R, D46S, D46T,D46V, D46W and D46Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 47 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 47 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprisesor consists of any of the substitutions selected from C47A, C47D, C47E,C47F, C47G, C47H, C47I, C47K, C47L, C47M, C47N, C47P, C47Q, C47R, C47S,C47T, C47V, C47W and C47Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 48 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 48 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Thr, Val or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from W48A, W48D, W48E,W48F, W48G, W48H, W48I, W48K, W48L, W48M, W48N, W48P, W48Q, W48R, W48S,W48T, W48V and W48Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 49 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 49 is substitutedwith Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from A49D, A49E, A49F,A49G, A49H, A49I, A49K, A49L, A49M, A49N, A49P, A49Q, A49R, A49S, A49T,A49V, A49W and A49Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 50 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 50 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from I50A, I50D, I50E,I50F, I50G, I50H, I50K, I50L, I50M, I50N, I50P, I50Q, I50R, I50S, I50T,I50V, I50W and I50Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 51 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 51 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from L51A, L51D, L51E,L51F, L51G, L51H, L51I, L51K, L51M, L51N, L51P, L51Q, L51R, L51S, L51T,L51V, L51W and L51Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution L51G ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 53 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 53 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from K53A, K53D, K53E,K53F, K53G, K53H, K53I, K53L, K53M, K53N, K53P, K53Q, K53R, K53S, K53T,K53V, K53W and K53Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsK53T or K53P of the polypeptide shown in SEQ ID NO: 1 or a polypeptidehaving at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 54 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 54 is substitutedwith Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from G54A, G54D, G54E,G54F, G54H, G54I, G54K, G54L, G54M, G54N, G54P, G54Q, G54R, G54S, G54T,G54V, G54W and G54Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution G54P ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 55 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 55 is substitutedwith Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from A55D, A55E, A55F,A55G, A55H, A55I, A55K, A55L, A55M, A55N, A55P, A55Q, A55R, A55S, A55T,A55V, A55W and A55Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution A55P ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 56 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 56 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from P56A, P56D, P56E,P56F, P56G, P56H, P56I, P56K, P56L, P56M, P56N, P56Q, P56R, P56S, P56T,P56V, P56W and P56Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 57 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 57 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from N57A, N57D, N57E,N57F, N57G, N57H, N57I, N57K, N57L, N57M, N57P, N57Q, N57R, N57S, N57T,N57V, N57W and N57Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution N57H ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 58 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 58 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Thr, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from V58A, V58D, V58E,V58F, V58G, V58H, V58I, V58K, V58L, V58M, V58N, V58P, V58Q, V58R, V58S,V58T, V58W and V58Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 59 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 59 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from L59A, L59D, L59E,L59F, L59G, L59H, L59I, L59K, L59M, L59N, L59P, L59Q, L59R, L59S, L59T,L59V, L59W and L59Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 60 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 60 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from Q60A, Q60D, Q60E,Q60F, Q60G, Q60H, Q60I, Q60K, Q60L, Q60M, Q60N, Q60P, Q60R, Q60S, Q60T,Q60V, Q60W and Q60Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 61 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 61 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from R61A, R61D, R61E,R61F, R61G, R61H, R61I, R61K, R61L, R61M, R61N, R61P, R61Q, R61S, R61T,R61V, R61W and R61Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 62 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 62 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Thr, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from V62A, V62D, V62E,V62F, V62G, V62H, V62I, V62K, V62L, V62M, V62N, V62P, V62Q, V62R, V62S,V62T, V62W and V62Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 63 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 63 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from N63A, N63D, N63E,N63F, N63G, N63H, N63I, N63K, N63L, N63M, N63P, N63Q, N63R, N63S, N63T,N63V, N63W and N63Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 64 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 64 is substitutedwith Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from E64A, E64D, E64F,E64G, E64H, E64I, E64K, E64L, E64M, E64N, E64P, E64Q, E64R, E64S, E64T,E64V, E64W and E64Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsE64A, E64Q, E64R, E64T, E641 or E64S of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 65 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 65 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from K65A, K65D, K65E,K65F, K65G, K65H, K65I, K65L, K65M, K65N, K65P, K65Q, K65R, K65S, K65T,K65V, K65W and K65Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 66 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 66 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from T66A, T66D, T66E,T66F, T66G, T66H, T66I, T66K, T66L, T66M, T66N, T66P, T66Q, T66R, T66S,T66V, T66W and T66Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitutions T66H ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 67 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 67 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from K67A, K67D, K67E,K67F, K67G, K67H, K67I, K67L, K67M, K67N, K67P, K67Q, K67R, K67S, K67T,K67V, K67W and K67Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsK67A or K67T of the polypeptide shown in SEQ ID NO: 1 or a polypeptidehaving at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 68 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 68 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from N68A, N68D, N68E,N68F, N68G, N68H, N68I, N68K, N68L, N68M, N68P, N68Q, N68R, N68S, N68T,N68V, N68W and N68Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsN68V, N68P, N681 or N68H of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 69 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 69 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from S69A, S69D, S69E,S69F, S69G, S69H, S69I, S69K, S69L, S69M, S69N, S69P, S69Q, S69R, S69T,S69V, S69W and S69Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsS69A, S69D, S69E, S69K, S69L, S69W, S69Y or S69Q of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 70 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 70 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from N70A, N70D, N70E,N70F, N70G, N70H, N70I, N70K, N70L, N70M, N70P, N70Q, N70R, N70S, N70T,N70V, N70W and N70Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsN70T, N70H or N70G of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 71 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 71 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from R71A, R71D, R71E,R71F, R71G, R71H, R71I, R71K, R71L, R71M, R71N, R71P, R71Q, R71S, R71T,R71V, R71W and R71Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution R71T ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 72 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 72 is substitutedwith Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from D72A, D72E, D72F,D72G, D72H, D72I, D72K, D72L, D72M, D72N, D72P, D72Q, D72R, D72S, D72T,D72V, D72W and D72Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution D72E ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 73 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 73 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Ser, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from R73A, R73D, R73E,R73F, R73G, R73H, R73I, R73K, R73L, R73M, R73N, R73P, R73Q, R73S, R73T,R73V, R73W and R73Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 74 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 74 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from S74A, S74D, S74E,S74F, S74G, S74H, S74I, S74K, S74L, S74M, S74N, S74P, S74Q, S74R, S74T,S74V, S74W and S74Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsS74H or S74G of the polypeptide shown in SEQ ID NO: 1 or a polypeptidehaving at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 75 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 75 is substitutedwith Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from G75A, G75D, G75E,G75F, G75H, G75I, G75K, G75L, G75M, G75N, G75P, G75Q, G75R, G75S, G75T,G75V, G75W and G75Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution G75I ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 76 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 76 is substitutedwith Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from A76D, A76E, A76F,A76G, A76H, A76I, A76K, A76L, A76M, A76N, A76P, A76Q, A76R, A76S, A76T,A76V, A76W and A76Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 77 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 77 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from N77A, N77D, N77E,N77F, N77G, N77H, N77I, N77K, N77L, N77M, N77P, N77Q, N77R, N77S, N77T,N77V, N77W and N77Y, of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitutions N77T ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 78 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 78 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from K78A, K78D, K78E,K78F, K78G, K78H, K78I, K78L, K78M, K78N, K78P, K78Q, K78R, K78S, K78T,K78V, K78W and K78Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 79 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 79 is substitutedwith Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from G79A, G79D, G79E,G79F, G79H, G79I, G79K, G79L, G79M, G79N, G79P, G79Q, G79R, G79S, G79T,G79V, G79W and G79Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 80 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 80 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from P80A, P80D, P80E,P80F, P80G, P80H, P80I, P80K, P80L, P80M, P80N, P80Q, P80R, P80S, P80T,P80V, P80W and P80Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 81 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 81 is substitutedwith Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from F81A, F81D, F81E,F81G, F81H, F81I, F81K, F81L, F81M, F81N, F81P, F81Q, F81R, F81S, F81T,F81V, F81W and F81Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 82 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 82 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from K82A, K82D, K82E,K82F, K82G, K82H, K82I, K82L, K82M, K82N, K82P, K82Q, K82R, K82S, K82T,K82V, K82W and K82Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsK82P or K82I of the polypeptide shown in SEQ ID NO: 1 or a polypeptidehaving at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 83 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 83 is substitutedwith Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from D83A, D83E, D83F,D83G, D83H, D83I, D83K, D83L, D83M, D83N, D83P, D83Q, D83R, D83S, D83T,D83V, D83W and D83Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsD83T, D83P, D83I, D83H or D83G of the polypeptide shown in SEQ ID NO: 1or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 84 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 84 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from P84A, P84D, P84E,P84F, P84G, P84H, P84I, P84K, P84L, P84M, P84N, P84Q, P84R, P84S, P84T,P84V, P84W and P84Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution P84H ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 85 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 85 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from Q85A, Q85D, Q85E,Q85F, Q85G, Q85H, Q85I, Q85K, Q85L, Q85M, Q85N, Q85P, Q85R, Q85S, Q85T,Q85V, Q85W and Q85Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsQ85T, Q85P or Q85H of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 86 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 86 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from K86A, K86D, K86E,K86F, K86G, K86H, K86I, K86L, K86M, K86N, K86P, K86Q, K86R, K86S, K86T,K86V, K86W and K86Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsK86T, K86P or K86H of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 87 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 87 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Ser, Arg, Thr, Val or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from W87A, W87D, W87E,W87F, W87G, W87H, W87I, W87K, W87L, W87M, W87N, W87P, W87Q, W87R, W87S,W87T, W87V and W87Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 88 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 88 is substitutedwith Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from G88A, G88D, G88E,G88F, G88H, G88I, G88K, G88L, G88M, G88N, G88P, G88Q, G88R, G88S, G88T,G88V, G88W and G88Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of any of the substitutionsG88P or G88H of the polypeptide shown in SEQ ID NO: 1 or a polypeptidehaving at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 89 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 89 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from I89A, I89D, I89E,I89F, I89G, I89H, I89K, I89L, I89M, I89N, I89P, I89Q, I89R, I89S, I89T,I89V, I89W and I89Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 90 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 90 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Ser, Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprisesor consists of any of the substitutions selected from K90A, K90D, K90E,K90F, K90G, K90H, K90I, K90L, K90M, K90N, K90P, K90Q, K90R, K90S, K90T,K90V, K90W and K90Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 91 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 91 is substitutedwith Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from A91D, A91E, A91F,A91G, A91H, A91I, A91K, A91L, A91M, A91N, A91P, A91Q, A91R, A91S, A91T,A91V, A91W and A91Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitution A91P ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 92 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 92 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from L92A, L92D, L92E,L92F, L92G, L92H, L92I, L92K, L92M, L92N, L92P, L92Q, L92R, L92S, L92T,L92V, L92W and L92Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 93 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 93 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from P93A, P93D, P93E,P93F, P93G, P93H, P93I, P93K, P93L, P93M, P93N, P93Q, P93R, P93S, P93T,P93V, P93W and P93Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 94 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 94 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from P94A, P94D, P94E,P94F, P94G, P94H, P94I, P94K, P94L, P94M, P94N, P94Q, P94R, P94S, P94T,P94V, P94W and P94Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 95 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 95 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from K95A, K95D, K95E,K95F, K95G, K95H, K95I, K95L, K95M, K95N, K95P, K95Q, K95R, K95S, K95T,K95V, K95W and K95Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 96 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 96 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from N96A, N96D, N96E,N96F, N96G, N96H, N96I, N96K, N96L, N96M, N96P, N96Q, N96R, N96S, N96T,N96V, N96W and N96Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 97 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 97 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln, Ser,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from P97A, P97D, P97E,P97F, P97G, P97H, P97I, P97K, P97L, P97M, P97N, P97Q, P97R, P97S, P97T,P97V, P97W and P97Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 98 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 98 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Thr, Val, Trp or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from S98A, S98D, S98E,S98F, S98G, S98H, S98I, S98K, S98L, S98M, S98N, S98P, S98Q, S98R, S98T,S98V, S98W and S98Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 99 of SEQ ID NO 1. In another aspect,the amino acid at a position corresponding to position 99 is substitutedwith Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Thr, Val or Tyr. In another aspect, the variant comprises orconsists of any of the substitutions selected from W99A, W99D, W99E,W99F, W99G, W99H, W99I, W99K, W99L, W99M, W99N, W99P, W99Q, W99R, W99S,W99T, W99V and W99Y of the polypeptide shown in SEQ ID NO: 1 or apolypeptide having at least 80% sequence identity hereto. In a preferredaspect, the variant comprises or consists of the substitutions W99T ofthe polypeptide shown in SEQ ID NO: 1 or a polypeptide having at least80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 100 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 100 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from S100A,S100D, S100E, S100F, S100G, S100H, S100I, S100K, S100L, S100M, S100N,S100P, S100Q, S100R, S100T, S100V, S100W and S100Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 101 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 101 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A101D,A101E, A101F, A101G, A101H, A101I, A101K, A101L, A101M, A101N, A101P,A101Q, A101R, A101S, A101T, A101V, A101W and A101Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution A101W of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 102 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 102 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from Q102A,Q102D, Q102E, Q102F, Q102G, Q102H, Q102I, Q102K, Q102L, Q102M, Q102N,Q102P, Q102R, Q102S, Q102T, Q102V, Q102W and Q102Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 103 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 103 issubstituted with Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Gln,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from D103A,D103E, D103F, D103G, D103H, D103I, D103K, D103L, D103M, D103N, D103P,D103Q, D103R, D103S, D103T, D103V, D103W and D103Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 104 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 104 issubstituted with Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Gln,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from F104A,F104D, F104E, F104G, F104H, F104I, F104K, F104L, F104M, F104N, F104P,F104Q, F104R, F104S, F104T, F104V, F104W and F104Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 105 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 105 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K105A,K105D, K105E, K105F, K105G, K105H, K105I, K105L, K105M, K105N, K105P,K105Q, K105R, K105S, K105T, K105V, K105W and K105Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of any of the substitutions K105E, K105N, K105T or K105D of thepolypeptide shown in SEQ ID NO: 1 or a polypeptide having at least 80%sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 106 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 106 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Gln, Pro, Arg, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from S106A,S106D, S106E, S106F, S106G, S106H, S106I, S106K, S106L, S106M, S106N,S106P, S106Q, S106R, S106T, S106V, S106W and S106Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution S106T of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 107 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 107 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from P107A,P107D, P107E, P107F, P107G, P107H, P107I, P107K, P107L, P107M, P107N,P107Q, P107R, P107S, P107T, P107V, P107W and P107Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 108 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 108 issubstituted with Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from E108A,E108D, E108F, E108G, E108H, E108I, E108K, E108L, E108M, E108N, E108P,E108Q, E108R, E108S, E108T, E108V, E108W and E108Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 109 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 109 issubstituted with Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from E109A,E109D, E109F, E109G, E109H, E109I, E109K, E109L, E109M, E109N, E109P,E109Q, E109R, E109S, E109T, E109V, E109W and E109Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 110 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 110 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Val or Trp. In another aspect, the variantcomprises or consists of any of the substitutions selected from Y110A,Y110D, Y110E, Y110F, Y110G, Y110H, Y110I, Y110K, Y110L, Y110M, Y110N,Y110P, Y110Q, Y110R, Y110S, Y110T, Y110V and Y110W of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 111 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 111 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A111D,A111E, A111F, A111G, A111H, A111I, A111K, A111L, A111M, A111N, A111P,A111Q, A111R, A111S, A111T, A111V, A111W and A111Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 112 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 112 issubstituted with Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from F112A,F112D, F112E, F112G, F112H, F112I, F112K, F112L, F112M, F112N, F112P,F112Q, F112R, F112S, F112T, F112V, F112W and F112Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 113 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 113 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A113D,A113E, A113F, A113G, A113H, A113I, A113K, A113L, A113M, A113N, A113P,A113Q, A113R, A113S, A113T, A113V, A113W and A113Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 114 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 114 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from S114A,S114D, S114E, S114F, S114G, S114H, S114I, S114K, S114L, S114M, S114N,S114P, S114Q, S114R, S114T, S114V, S114W and S114Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 115 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 115 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from S115A,S115D, S115E, S115F, S115G, S115H, S115I, S115K, S115L, S115M, S115N,S115P, S115Q, S115R, S115T, S115V, S115W and S115Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution S115T of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 116 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 116 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from L116A,L116D, L116E, L116F, L116G, L116H, L116I, L116K, L116M, L116N, L116P,L116Q, L116R, L116S, L116T, L116V, L116W and L116Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution L116I of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 117 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 117 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from Q117A,Q117D, Q117E, Q117F, Q117G, Q117H, Q117I, Q117K, Q117L, Q117M, Q117N,Q117P, Q117R, Q117S, Q117T, Q117V, Q117W and Q117Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 118 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 118 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G118A,G118D, G118E, G118F, G118H, G118I, G118K, G118L, G118M, G118N, G118P,G118Q, G118R, G118S, G118T, G118V, G118W and G118Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 119 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 119 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G119A,G119D, G119E, G119F, G119H, G119I, G119K, G119L, G119M, G119N, G119P,G119Q, G119R, G119S, G119T, G119V, G119W and G119Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 120 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 120 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from T120A,T120D, T120E, T120F, T120G, T120H, T120I, T120K, T120L, T120M, T120N,T120P, T120Q, T120R, T120S, T120V, T120W and T120Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 121 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 121 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from N121A,N121D, N121E, N121F, N121G, N121H, N121I, N121I, N121K, N121L, N121M,N121P, N121Q, N121R, N121S, N121T, N121V, N121W and N121Y of thepolypeptide shown in SEQ ID NO: 1 or a polypeptide having at least 80%sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 122 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 122 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A122D,A122E, A122F, A122G, A122H, A122I, A122K, A122L, A122M, A122N, A122P,A122Q, A122R, A122S, A122T, A122V, A122W and A122Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 123 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 123 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from I123A,I123D, I123E, I123F, I123G, I123H, I123K, I123L, I123M, I123N, I123P,I123Q, I123R, I123S, I123T, I123V, I123W and I123Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 124 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 124 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from L124A,L124D, L124E, L124F, L124G, L124H, L124I, L124K, L124M, L124N, L124P,L124Q, L124R, L124S, L124T, L124V, L124W and L124Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 125 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 125 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A125D,A125E, A125F, A125G, A125H, A125I, A125K, A125L, A125M, A125N, A125P,A125Q, A125R, A125S, A125T, A125V, A125W and A125Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 126 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 126 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from P126A,P126D, P126E, P126F, P126G, P126H, P126I, P126K, P126L, P126M, P126N,P126Q, P126R, P126S, P126T, P126V, P126W and P126Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 127 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 127 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from V127A,V127D, V127E, V127F, V127G, V127H, V127I, V127K, V127L, V127M, V127N,V127P, V127Q, V127R, V127S, V127T, V127W and V127Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 128 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 128 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from N128A,N128D, N128E, N128F, N128G, N128H, N128I, N128K, N128L, N128M, N128P,N128Q, N128R, N128S, N128T, N128V, N128W and N128Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 129 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 129 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from L129A,L129D, L129E, L129F, L129G, L129H, L129I, L129K, L129M, L129N, L129P,L129Q, L129R, L129S, L129T, L129V, L129W and L129Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 130 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 130 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A130D,A130E, A130F, A130G, A130H, A130I, A130K, A130L, A130M, A130N, A130P,A130Q, A130R, A130S, A130T, A130V, A130W and A130Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 131 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 131 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from S131A,S131D, S131E, S131F, S131G, S131H, S131I, S131K, S131L, S131M, S131N,S131P, S131Q, S131R, S131T, S131V, S131W and S131Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 132 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 132 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from Q132A,Q132D, Q132E, Q132F, Q132G, Q132H, Q132I, Q132K, Q132L, Q132M, Q132N,Q132P, Q132R, Q132S, Q132T, Q132V, Q132W and Q132Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 133 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 133 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from N133A,N133D, N133E, N133F, N133G, N133H, N133I, N133K, N133L, N133M, N133P,N133Q, N133R, N133S, N133T, N133V, N133W or N133Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 134 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 134 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from S134A,S134D, S134E, S134F, S134G, S134H, S134I, S134K, S134L, S134M, S134N,S134P, S134Q, S134R, S134T, S134V, S134W and S134Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 135 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 135 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from Q135A,Q135D, Q135E, Q135F, Q135G, Q135H, Q135I, Q135K, Q135L, Q135M, Q135N,Q135P, Q135R, Q135S, Q135T, Q135V, Q135W and Q135Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution Q135L of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 136 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 136 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G136A,G136D, G136E, G136F, G136H, G136I, G136K, G136L, G136M, G136N, G136P,G136Q, G136R, G136S, G136T, G136V, G136W and G136Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution G136L of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 137 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 137 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G137A,G137D, G137E, G137F, G137H, G137I, G137K, G137L, G137M, G137N, G137P,G137Q, G137R, G137S, G137T, G137V, G137W and G137Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 138 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 138 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from V138A,V138D, V138E, V138F, V138G, V138H, V138I, V138K, V138L, V138M, V138N,V138P, V138Q, V138R, V138S, V138T, V138W and V138Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of any of the substitutions V138I, V138L, V138P or V138Q of thepolypeptide shown in SEQ ID NO: 1 or a polypeptide having at least 80%sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 139 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 139 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from L139A,L139D, L139E, L139F, L139G, L139H, L139I, L139K, L139M, L139N, L139P,L139Q, L139R, L139S, L139T, L139V, L139W and L139Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution L139A of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 140 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 140 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from N140A,N140D, N140E, N140F, N140G, N140H, N140I, N140K, N140L, N140M, N140P,N140Q, N140R, N140S, N140T, N140V, N140W and N140Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of any of the substitutions N140R, N140L or N140A of thepolypeptide shown in SEQ ID NO: 1 or a polypeptide having at least 80%sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 141 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 141 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G141A,G141D, G141E, G141F, G141H, G141I, G141K, G141L, G141M, G141N, G141P,G141Q, G141R, G141S, G141T, G141V, G141W and G141Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution G141L of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 142 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 142 issubstituted with Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from F142A,F142D, F142E, F142G, F142H, F142I, F142K, F142L, F142M, F142N, F142P,F142Q, F142R, F142S, F142T, F142V, F142W and F142Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 143 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 143 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Val or Trp. In another aspect, the variantcomprises or consists of any of the substitutions selected from Y143A,Y143D, Y143E, Y143F, Y143G, Y143H, Y143I, Y143K, Y143L, Y143M, Y143N,Y143P, Y143Q, Y143R, Y143S, Y143T, Y143V and Y143W of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 144 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 144 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from S144A,S144D, S144E, S144F, S144G, S144H, S144I, S144K, S144L, S144M, S144N,S144P, S144Q, S144R, S144T, S144V, S144W and S144Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 145 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 145 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A145D,A145E, A145F, A145G, A145H, A145I, A145K, A145L, A145M, A145N, A145P,A145Q, A145R, A145S, A145T, A145V, A145W and A145Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 146 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 146 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from N146A,N146D, N146E, N146F, N146G, N146H, N146I, N146K, N146L, N146M, N146P,N146Q, N146R, N146S, N146T, N146V, N146W and N146Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 147 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 147 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K147A,K147D, K147E, K147F, K147G, K147H, K147I, K147L, K147M, K147N, K147P,K147Q, K147R, K147S, K147T, K147V, K147W and K147Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 148 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 148 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from V148A,V148D, V148E, V148F, V148G, V148H, V148I, V148K, V148L, V148M, V148N,V148P, V148Q, V148R, V148S, V148T, V148W and V148Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 149 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 149 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A149D,A149E, A149F, A149G, A149H, A149I, A149K, A149L, A149M, A149N, A149P,A149Q, A149R, A149S, A149T, A149V, A149W and A149Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 150 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 150 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from Q150A,Q150D, Q150E, Q150F, Q150G, Q150H, Q150I, Q150K, Q150L, Q150M, Q150N,Q150P, Q150R, Q150S, Q150T, Q150V, Q150W and Q150Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 151 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 151 issubstituted with Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from F151A,F151D, F151E, F151G, F151H, F151I, F151K, F151L, F151M, F151N, F151P,F151Q, F151R, F151S, F151T, F151V, F151W and F151Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution F151R of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 152 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 152 issubstituted with Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from D152A,D152E, D152F, D152G, D152H, D152I, D152K, D152L, D152M, D152N, D152P,D152Q, D152R, D152S, D152T, D152V, D152W and D152Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of any of the substitutions D152Y, D152L, D1521 or D152A of thepolypeptide shown in SEQ ID NO: 1 or a polypeptide having at least 80%sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 153 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 153 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from P153A,P153D, P153E, P153F, P153G, P153H, P153I, P153K, P153L, P153M, P153N,P153Q, P153R, P153S, P153T, P153V, P153W and P153Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution P153E of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 154 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 154 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from S154A,S154D, S154E, S154F, S154G, S154H, S154I, S154K, S154L, S154M, S154N,S154P, S154Q, S154R, S154T, S154V, S154W and S154Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution S154R of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 155 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 155 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K155A,K155D, K155E, K155F, K155G, K155H, K155I, K155L, K155M, K155N, K155P,K155Q, K155R, K155S, K155T, K155V, K155W and K155Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 156 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 156 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from P156A,P156D, P156E, P156F, P156G, P156H, P156I, P156K, P156L, P156M, P156N,P156Q, P156R, P156S, P156T, P156V, P156W and P156Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 157 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 157 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from Q157A,Q157D, Q157E, Q157F, Q157G, Q157H, Q157I, Q157K, Q157L, Q157M, Q157N,Q157P, Q157R, Q157S, Q157T, Q157V, Q157W and Q157Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 158 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 158 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from Q158A,Q158D, Q158E, Q158F, Q158G, Q158H, Q158I, Q158K, Q158L, Q158M, Q158N,Q158P, Q158R, Q158S, Q158T, Q158V, Q158W and Q158Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 159 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 159 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from T159A,T159D, T159E, T159F, T159G, T159H, T159I, T159K, T159L, T159M, T159N,T159P, T159Q, T159R, T159S, T159V, T159W and T159Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 160 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 160 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K160A,K160D, K160E, K160F, K160G, K160H, K160I, K160L, K160M, K160N, K160P,K160Q, K160R, K160S, K160T, K160V, K160W and K160Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 161 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 161 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G161A,G161D, G161E, G161F, G161H, G161I, G161K, G161L, G161M, G161N, G161P,G161Q, G161R, G161S, G161T, G161V, G161W and G161Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 162 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 162 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from T162A,T162D, T162E, T162F, T162G, T162H, T162I, T162K, T162L, T162M, T162N,T162P, T162Q, T162R, T162S, T162T, T162V, T162W and T162Y of thepolypeptide shown in SEQ ID NO: 1 or a polypeptide having at least 80%sequence identity hereto. In a preferred aspect, the variant comprisesor consists of the substitution T162R of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 163 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 163 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Val or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from W163A,W163D, W163E, W163F, W163G, W163H, W163I, W163K, W163L, W163M, W163N,W163P, W163Q, W163R, W163S, W163T, W163V and W163Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution W163E of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 164 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 164 issubstituted with Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from F164A,F164D, F164E, F164G, F164H, F164I, F164K, F164L, F164M, F164N, F164P,F164Q, F164R, F164S, F164T, F164V, F164W and F164Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution F164R of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 165 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 165 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from Q165A,Q165D, Q165E, Q165F, Q165G, Q165H, Q165I, Q165K, Q165L, Q165M, Q165N,Q165P, Q165R, Q165S, Q165T, Q165V, Q165W and Q165Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 166 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 166 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from I166A,I166D, I166E, I166F, I166G, I166H, I166K, I166L, I166M, I166N, I166P,I166Q, I166R, I166S, I166T, I166V, I166W and I166Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of any of the substitutions I166Y or I166R of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 167 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 167 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from T167A,T167D, T167E, T167F, T167G, T167H, T167I, T167K, T167L, T167M, T167N,T167P, T167Q, T167R, T167S, T167V, T167W and T167Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 168 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 168 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K168A,K168D, K168E, K168F, K168G, K168H, K168I, K168L, K168M, K168N, K168P,K168Q, K168R, K168S, K168T, K168V, K168W and K168Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution K168N of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 169 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 169 issubstituted with Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from F169A,F169D, F169E, F169G, F169H, F169I, F169K, F169L, F169M, F169N, F169P,F169Q, F169R, F169S, F169T, F169V, F169W and F169Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of any of the substitutions F169R or F169E of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 170 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 170 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from T170A,T170D, T170E, T170F, T170G, T170H, T170I, T170K, T170L, T170M, T170N,T170P, T170Q, T170R, T170S, T170V, T170W and T170Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 171 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 171 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G171A,G171D, G171E, G171F, G171H, G171I, G171K, G171L, G171M, G171N, G171P,G171Q, G171R, G171S, G171T, G171V, G171W and G171Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 172 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 172 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A172D,A172E, A172F, A172G, A172H, A172I, A172K, A172L, A172M, A172N, A172P,A172Q, A172R, A172S, A172T, A172V, A172W and A172Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 173 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 173 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A173D,A173E, A173F, A173G, A173H, A173I, A173K, A173L, A173M, A173N, A173P,A173Q, A173R, A173S, A173T, A173V, A173W and A173Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of any of the substitutions A173I, A173R or A173T of thepolypeptide shown in SEQ ID NO: 1 or a polypeptide having at least 80%sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 174 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 174 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G174A,G174D, G174E, G174F, G174H, G174I, G174K, G174L, G174M, G174N, G174P,G174Q, G174R, G174S, G174T, G174V, G174W and G174Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 175 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 175 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from P175A,P175D, P175E, P175F, P175G, P175H, P175I, P175K, P175L, P175M, P175N,P175Q, P175R, P175S, P175T, P175V, P175W and P175Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 176 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 176 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Val or Trp. In another aspect, the variantcomprises or consists of any of the substitutions selected from Y176A,Y176D, Y176E, Y176F, Y176G, Y176H, Y176I, Y176K, Y176L, Y176M, Y176N,Y176P, Y176Q, Y176R, Y176S, Y176T, Y176V and Y176W of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 178 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 178 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K178A,K178D, K178E, K178F, K178G, K178H, K178I, K178L, K178M, K178N, K178P,K178Q, K178R, K178S, K178T, K178V, K178W and K178Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 179 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 179 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A179D,A179E, A179F, A179G, A179H, A179I, A179K, A179L, A179M, A179N, A179P,A179Q, A179R, A179S, A179T, A179V, A179W and A179Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 180 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 180 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from L180A,L180D, L180E, L180F, L180G, L180H, L180I, L180K, L180M, L180N, L180P,L180Q, L180R, L180S, L180T, L180V, L180W and L180Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 181 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 181 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G181A,G181D, G181E, G181F, G181H, G181I, G181K, G181L, G181M, G181N, G181P,G181Q, G181R, G181S, G181T, G181V, G181W and G181Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 182 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 182 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from S182A,S182D, S182E, S182F, S182G, S182H, S182I, S182K, S182L, S182M, S182N,S182P, S182Q, S182R, S182T, S182V, S182W and S182Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of any of the substitutions S182Y of the polypeptide shown inSEQ ID NO: 1 or a polypeptide having at least 80% sequence identityhereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 183 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 183 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from N183A,N183D, N183E, N183F, N183G, N183H, N183I, N183K, N183L, N183M, N183P,N183Q, N183R, N183S, N183T, N183V, N183W and N183Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution N183E of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 184 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 184 issubstituted with Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from D184A,D184E, D184F, D184G, D184H, D184I, D184K, D184L, D184M, D184N, D184P,D184Q, D184R, D184S, D184T, D184V, D184W and D184Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution D184I of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 185 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 185 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K185A,K185D, K185E, K185F, K185G, K185H, K185I, K185L, K185M, K185N, K185P,K185Q, K185R, K185S, K185T, K185V, K185W and K185Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution K185Y of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 186 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 186 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from S186A,S186D, S186E, S186F, S186G, S186H, S186I, S186K, S186L, S186M, S186N,S186P, S186Q, S186R, S186T, S186V, S186W and S186Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution S186I of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 187 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 187 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from V187A,V187D, V187E, V187F, V187G, V187H, V187I, V187K, V187L, V187M, V187N,V187P, V187Q, V187R, V187S, V187T, V187W and V187Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 189 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 189 issubstituted with Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from D189A,D189E, D189F, D189G, D189H, D189I, D189K, D189L, D189M, D189N, D189P,D189Q, D189R, D189S, D189T, D189V, D189W and D189Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of any of the substitutions D189G or D189H of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 190 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 190 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K190A,K190D, K190E, K190F, K190G, K190H, K190I, K190L, K190M, K190N, K190P,K190Q, K190R, K190S, K190T, K190V, K190W and K190Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 191 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 191 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from N191A,N191D, N191E, N191F, N191G, N191H, N191I, N191K, N191L, N191M, N191P,N191Q, N191R, N191S, N191T, N191V, N191W and N191Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 192 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 192 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K192A,K192D, K192E, K192F, K192G, K192H, K192I, K192L, K192M, K192N, K192P,K192Q, K192R, K192S, K192T, K192V, K192W and K192Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 193 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 193 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from N193A,N193D, N193E, N193F, N193G, N193H, N193I, N193K, N193L, N193M, N193P,N193Q, N193R, N193S, N193T, N193V, N193W and N193Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 194 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 194 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from I194A,I194D, I194E, I194F, I194G, I194H, I194K, I194L, I194M, I194N, I194P,I194Q, I194R, I194S, I194T, I194V, I194W and I194Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 195 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 195 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A195D,A195E, A195F, A195G, A195H, A195I, A195K, A195L, A195M, A195N, A195P,A195Q, A195R, A195S, A195T, A195V, A195W and A195Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 196 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 196 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G196A,G196D, G196E, G196F, G196H, G196I, G196K, G196L, G196M, G196N, G196P,G196Q, G196R, G196S, G196T, G196V, G196W and G196Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 197 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 197 issubstituted with Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from D197A,D197E, D197F, D197G, D197H, D197I, D197K, D197L, D197M, D197N, D197P,D197Q, D197R, D197S, D197T, D197V, D197W and D197Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 198 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 198 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Val, or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from W198A,W198D, W198E, W198F, W198G, W198H, W198I, W198K, W198L, W198M, W198N,W198P, W198Q, W198R, W198S, W198T, W198V and W198Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 199 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 199 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G199A,G199D, G199E, G199F, G199H, G199I, G199K, G199L, G199M, G199N, G199P,G199Q, G199R, G199S, G199T, G199V, G199W and G199Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 200 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 200 issubstituted with Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from F200A,F200D, F200E, F200G, F200H, F200I, F200K, F200L, F200M, F200N, F200P,F200Q, F200R, F200S, F200T, F200V, F200W and F200Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 201 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 201 issubstituted with Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from D201A,D201E, D201F, D201G, D201H, D201I, D201K, D201L, D201M, D201N, D201P,D201Q, D201R, D201S, D201T, D201V, D201W and D201Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 202 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 202 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from P202A,P202D, P202E, P202F, P202G, P202H, P202I, P202K, P202L, P202M, P202N,P202Q, P202R, P202S, P202T, P202V, P202W and P202Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 203 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 203 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A203D,A203E, A203F, A203G, A203H, A203I, A203K, A203L, A203M, A203N, A203P,A203Q, A203R, A203S, A203T, A203V, A203W and A203Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 204 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 204 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K204A,K204D, K204E, K204F, K204G, K204H, K204I, K204L, K204M, K204N, K204P,K204Q, K204R, K204S, K204T, K204V, K204W and K204Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 205 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 205 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Val or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from W205A,W205D, W205E, W205F, W205G, W205H, W205I, W205K, W205L, W205M, W205N,W205P, W205Q, W205R, W205S, W205T, W205V and W205Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 206 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 206 issubstituted with Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from A206D,A206E, A206F, A206G, A206H, A206I, A206K, A206L, A206M, A206N, A206P,A206Q, A206R, A206S, A206T, A206V, A206W and A206Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 207 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 207 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Val or Trp. In another aspect, the variantcomprises or consists of any of the substitutions selected from Y207A,Y207D, Y207E, Y207F, Y207G, Y207H, Y207I, Y207K, Y207L, Y207M, Y207N,Y207P, Y207Q, Y207R, Y207S, Y207T, Y207V and Y207W of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 208 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 208 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from Q208A,Q208D, Q208E, Q208F, Q208G, Q208H, Q208I, Q208K, Q208L, Q208M, Q208N,Q208P, Q208R, Q208S, Q208T, Q208V, Q208W and Q208Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 209 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 209 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Val or Trp. In another aspect, the variantcomprises or consists of any of the substitutions selected from Y209A,Y209D, Y209E, Y209F, Y209G, Y209H, Y209I, Y209K, Y209L, Y209M, Y209N,Y209P, Y209Q, Y209R, Y209S, Y209T, Y209V and Y209W of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 210 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 210 issubstituted with Ala, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from D210A,D210E, D210F, D210G, D210H, D210I, D210K, D210L, D210M, D210N, D210P,D210Q, D210R, D210S, D210T, D210V, D210W and D210Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 211 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 211 issubstituted with Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from E211A,E211D, E211F, E211G, E211H, E211I, E211K, E211L, E211M, E211N, E211P,E211Q, E211R, E211S, E211T, E211V, E211W and E211Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 212 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 212 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K212A,K212D, K212E, K212F, K212G, K212H, K212I, K212L, K212M, K212N, K212P,K212Q, K212R, K212S, K212T, K212V, K212W and K212Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of any of the substitutions K212G or K212P of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 213 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 213 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from N213A,N213D, N213E, N213F, N213G, N213H, N213I, N213K, N213L, N213M, N213P,N213Q, N213R, N213S, N213T, N213V, N213W and N213Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 214 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 214 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from N214A,N214D, N214E, N214F, N214G, N214H, N214I, N214K, N214L, N214M, N214P,N214Q, N214R, N214S, N214T, N214V, N214W and N214Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 215 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 215 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K215A,K215D, K215E, K215F, K215G, K215H, K215I, K215L, K215M, K215N, K215P,K215Q, K215R, K215S, K215T, K215V, K215W and K215Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto. In a preferred aspect, the variant comprises orconsists of the substitution K215I of the polypeptide shown in SEQ IDNO: 1 or a polypeptide having at least 80% sequence identity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 216 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 216 issubstituted with Ala, Asp, Glu, Gly, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from F216A,F216D, F216E, F216G, F216H, F216I, F216K, F216L, F216M, F216N, F216P,F216Q, F216R, F216S, F216T, F216V, F216W and F216Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 217 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 217 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from N217A,N217D, N217E, N217F, N217G, N217H, N217I, N217K, N217L, N217M, N217P,N217Q, N217R, N217S, N217T, N217V, N217W and N217Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 218 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 218 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Val or Trp. In another aspect, the variantcomprises or consists of any of the substitutions selected from Y218A,Y218D, Y218E, Y218F, Y218G, Y218H, Y218I, Y218K, Y218L, Y218M, Y218N,Y218P, Y218Q, Y218R, Y218S, Y218T, Y218V and Y218W of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 219 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 219 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn,Pro, Gln, Arg, Ser, Thr, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from V219A,V219D, V219E, V219F, V219G, V219H, V219I, V219K, V219L, V219M, V219N,V219P, V219Q, V219R, V219S, V219T, V219W and V219Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 220 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 220 issubstituted with Ala, Asp, Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from G220A,G220D, G220E, G220F, G220H, G220I, G220K, G220L, G220M, G220N, G220P,G220Q, G220R, G220S, G220T, G220V, G220W and G220Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

In some aspect, the variant comprises or consists of an alteration at aposition corresponding to position 221 of SEQ ID NO 1. In anotheraspect, the amino acid at a position corresponding to position 221 issubstituted with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Pro,Gln, Arg, Ser, Thr, Val, Trp or Tyr. In another aspect, the variantcomprises or consists of any of the substitutions selected from K221A,K221D, K221E, K221F, K221G, K221H, K221I, K221L, K221M, K221N, K221P,K221Q, K221R, K221S, K221T, K221V, K221W and K221Y of the polypeptideshown in SEQ ID NO: 1 or a polypeptide having at least 80% sequenceidentity hereto.

By the term “of the polypeptide” shown in SEQ ID NO: 1 is meant that thesubstitutions is made in SEQ ID NO 1 and the positions correspond to thepositions of SEQ ID NO 1. Thus the above variants are variants of aDNase having SEQ ID NO 1 or a DNase having at least 80% sequenceidentity hereto. The above mentioned variants have at least at least60%, at least 61%, at least 62%, at least 63%, at least 64%, at least65%, at least 66%, at least 67%, at least 68%, at least 69%, at least70%, at least 72%, at least 73%, at least 74%, at least 75%, at least80%, at least 81%, at least 82%, at least 83%, at least 84%, at least85%, at least 90%, at least 91%, at least 92%, at least 93%, at least94%, at least 95%, at least 96%, at least 97%, at least 98%, at least99%, e. g. at least 99.1%, at least 99.2%, at least 99.3%, at least99.4%, at least 99.5 but less than 100% identity to a polypeptide shownin SEQ ID NO: 1.

Some aspect of the invention relates to a DNase variant wherein thevariant has DNase activity, wherein the DNase variant has at least 80%sequence identity to the polypeptide shown in SEQ ID NO 1 and whereinthe variant when compared to the polypeptide shown in SEQ ID NO 1comprises a deletion of the amino acid in any of the positions 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 48, 49, 50, 51, 53, 54, 55, 56, 57, 58, 59, 60,61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78,79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96,97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111,112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125,126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139,140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153,154, 155, 156, 157, 158, 159, 160, 161, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 178, 179, 180, 181, 182, 183,185, 186, 187, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199,200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213,214, 215, 216, 217, 218, 219, 220 or 221 of the polypeptide shown in SEQID NO: 1, with the proviso that maximum number of deletions are 10 aminoacids compared to the mature polypeptide shown in SEQ ID NO 1. Somepreferred aspect of the invention relates to a DNase variant wherein thevariant has DNase activity, wherein the DNase variant has at least 80%sequence identity to the polypeptide shown in SEQ ID NO 1 and whereinthe variant when compared to the polypeptide shown in SEQ ID NO 1comprises a deletion of the amino acid in any of the positions 84, 88,139, 179 or 180 of the polypeptide shown in SEQ ID NO: 1.

When the DNase variant of the invention comprises one or more deletionthe DNase variant preferably comprises 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10deletion(s) compared to the polypeptide shown in SEQ ID NO 1,preferably, 1, preferably 2, preferably 3 or preferably 4 deletion(s)compared to the polypeptide shown in SEQ ID NO 1, wherein the DNasevariant has DNase activity and wherein the DNase variant has at least atleast 60%, at least 61%, at least 62%, at least 63%, at least 64%, atleast 65%, at least 66%, at least 67%, at least 68%, at least 69%, atleast 70%, at least 72%, at least 73%, at least 74%, at least 75%, atleast 80%, at least 81%, at least 82%, at least 83%, at least 84%, atleast 85%, at least 90%, at least 91%, at least 92%, at least 93%, atleast 94%, at least 95%, at least 96%, at least 97%, at least 98%, atleast 99% but less than 100% identity to a polypeptide shown in SEQ IDNO: 1.

In one embodiment, the DNase variants of the invention have improvedDNase activity compared to a reference DNase e.g. SEQ ID NO: 1, whereinthe activity is tested in Assay I described in Assays and detergentcompositions below.

In one embodiment, the DNase variants of the invention have improvedstability in detergent compared to a reference DNase e.g. SEQ ID NO: 1,wherein stability is tested as described in example 2.

In one embodiment, the DNase variants generated in the library asdescribed above have improved deep cleaning performance compared to areference DNase, e.g. SEQ ID NO: 1.

In one embodiment, the DNase variants of the invention have improveddeep cleaning performance compared to a reference DNase, e.g. SEQ ID NO:1.

When items like T-shirts or sportswear are used, they are exposed tobacteria from the body of the user and from the rest of the environmentin which they are used. This may cause malodor on the item even afterthe item is washed. The present invention therefore also relates toremoval or reduction of malodor on textile. The malodor may be caused bybacteria producing compounds with an unpleasant smell. One example ofsuch unpleasant smelling compounds is E-2-nonenal. The malodor can bepresent on newly washed textile which is still wet. Or the malodor canbe present on newly washed textile, which has subsequently been dried.The malodor may also be present on textile, which has been stored forsome time after wash. The present invention may relates to the reductionor removal of malodor such as E-2-nonenal from wet or dry textile usinga DNase variant according to the invention.

In one embodiment, the DNase variants according to the invention haveimproved malodor removal properties compared to a reference DNase e.g.SEQ ID NO: 1, wherein the malodor is measured as described in Assay II.

Further, the invention relates to the use of a DNase variant accordingto the invention for preventing, reducing or removing the adherence ofsoil to an item. In one embodiment, the item is textile. When the soildoes not adhere to the item, the item appears cleaner. Thus, theinvention further concerns the use of a DNase variant according to theinvention for maintaining or improving the whiteness of the item.

The present invention further concerns detergent compositions comprisinga DNase variant according to the invention preferably with a detergentadjunct ingredient. The detergent composition comprising a DNasevariants according to the invention may be used for preventing, reducingor removing biofilm from an item, for preventing, reducing or removingthe stickiness of an item, for pretreating stains on the item, forpreventing, reducing or removing redeposition of soil during a washcycle, for reducing or removing adherence of soil to an item, formaintaining or improving the whiteness of an item and for preventing,reducing or removing malodor from an item, such as E-2-nonenal (asdescribed in Assay II).

In one embodiment, the detergent composition is a liquid or powderlaundry detergent, suitable for e.g. washing at high temperature and/orpH, such as at or above 400C and/or at or above pH 8. In one embodimentthe detergent composition is a liquid or powder laundry detergent,suitable for e.g. washing at low temperature and/or pH, such as at orbelow 20° C. and/or pH 6. The detergent may also be formulated as a unitdose detergent and/or compact detergent optionally with minimum or nowater. The detergent may also be a dish wash detergent. The laundry anddish wash detergents are preferably phosphate-free. The detergentcomposition may further comprise at least one additional enzyme, such ascarbohydrate-active enzymes like carbohydrase, pectinase, mannanase,amylase, cellulase, arabinase, galactanase, xylanase, protease such asmetalloproteases, lipase, a, cutinase, oxidase, e.g., a laccase, and/orperoxidase.

In some further embodiments, the present invention relates to DNasevariants having at least 60% to SEQ ID NO: 1 identity hereto whereinwhen the variant having at least one improved property compared to SEQID NO: 1 when tested in a relevant assay. One embodiment of theinvention relates to DNase variants having an improvement factor above 1when the DNase variants are tested for a property of interest in arelevant assay, wherein the property of the reference DNase is given avalue of 1. In one embodiment, the property is stability, such asstorage stability in detergent in another embodiment the property iswash performance, such as deep cleaning performance.

In one embodiment, the variants according to the invention have one ormore improved property relative to the parent measured as an ImprovementFactor (IF) that is greater than 1.0, wherein the improved property isstability such as stability in detergent.

In one embodiment, the variants according to the invention have anImprovement Factor (IF) which is greater than 1.0 in at least one of twodifferent set of conditions A or B as described in example 2.

In one embodiment, the variants according to the invention have anImprovement Factor (IF) which is greater than 1.0 in one or both ofconditions A and B.

In some aspects, the variants according to the invention have anImprovement Factor (IF) which is at least 1.1; 1.2; 1.3; 1.4; 1.5; 1.6;1.7; 1.8; 1.9; 2.0; 2.1; 2.3; 2.4; 2.5, 2.6, 2.7, 2.8, 2.9 or 3.0.

In some aspects, the variants according to the invention have anImprovement Factor (IF) which is at least 1.1; 1.5; 2.0; 3.0, 3.5, 4.0,5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5 or 10.

Amino acid positions within a protein that are useful for makingvariants are those positions wherein at least one alterations leads to avariant exhibiting an improved characteristic as compared to theunchanged protein i.e. parent i.e. IF>1.0. The improved characteristicmay be determined using the assay I or II or as described in example 2.

One embodiment of the invention relates to DNase variants having atleast 60% identity to SEQ ID NO: 1, having DNase activity and comprisingone or more alterations selected from the group consisting of V1*, V1A,V1D, V1E, V1F, V1G, V1H, V1I, V1K, V1L, V1M, V1N, V1P, V1Q, V1R, V1S,V1T, V1W, V1Y, P2*, P2A, P2D, P2E, P2F, P2G, P2H, P2I, P2K, P2L, P2M,P2N, P2Q, P2R, P2S, P2T, P2V, P2W, P2Y, V3*, V3A, V3C, V3D, V3E, V3F,V3G, V3H, V3I, V3K, V3L, V3M, V3N, V3P, V3R, V3S, V3T, V3W, V3Y, N4*,N4A, N4D, N4E, N4F, N4G, N4H, N4I, N4K, N4L, N4M, N4P, N4Q, N4R, N4S,N4T, N4V, N4W, N4Y, P5*, P5A, P5D, P5E, P5F, P5G, P5H, PSI, P5K, P5L,P5M, P5N, P5Q, P5R, P5S, P5T, P5V, P5W, P5Y, E6*, E6A, E6D, E6F, E6G,E6H, E6I, E6K, E6L, E6M, E6N, E6P, E6Q, E6R, E6S, E6T, E6V, E6W, E6Y,P7*, P7A, P7D, P7E, P7F, P7G, P7H, P7I, P7K, P7L, P7M, P7N, P7Q, P7R,P7S, P7T, P7V, P7W, P7Y, D8*, D8A, D8E, D8F, D8G, D8H, D8I, D8K, D8L,D8M, D8N, D8P, D8Q, D8R, D8S, D8T, D8V, D8W, D8Y, A9*, A9D, A9E, A9F,A9G, A9H, A9I, A9K, A9L, A9M, A9N, A9P, A9Q, A9R, A9S, A9T, A9V, A9W,A9Y, T10*, T10A, T10D, T10E, T10F, T10G, T10H, T10I, T10K, T10L, T10M,T10N, T10P, T10Q, T10R, T10S, T10V, T10W, T10Y, S11*, S11A, S11D, S11E,S11F, S11G, S11H, S11I, S11K, S11L, S11M, S11N, S11P, S11Q, S11R, S11T,S11V, S11W, S11Y, V12*, V12A, V12D, V12E, V12F, V12G, V12H, V12I, V12K,V12L, V12M, V12N, V12P, V12Q, V12R, V12S, V12T, V12W, V12Y, E13*, E13A,E13D, E13F, E13G, E13H, E13I, E13K, E13L, E13M, E13N, E13P, E13Q, E13R,E13S, E13T, E13V, E13W, E13Y, N14*, N14A, N14D, N14E, N14F, N14G, N14H,N14I, N14K, N14L, N14M, N14P, N14Q, N14R, N14S, N14T, N14V, N14W, N14Y,V15*, V15A, V15D, V15E, V15F, V15G, V15H, V15I, V15K, V15L, V15M, V15N,V15P, V15Q, V15R, V15S, V15T, V15W, V15Y, A16*, A16D, A16E, A16F, A16G,A16H, A16I, A16K, A16L, A16M, A16N, A16P, A16Q, A16R, A16S, A16T, A16V,A16W, A16Y, L17*, L17A, L17D, L17E, L17F, L17G, L17H, L17I, L17K, L17M,L17N, L17P, L17Q, L17R, L17S, L17T, L17V, L17W, L17Y, K18*, K18A, K18D,K18E, K18F, K18G, K18H, K18I, K18L, K18M, K18N, K18P, K18Q, K18R, K18S,K18T, K18V, K18W, K18Y, T19*, T19A, T19D, T19E, T19F, T19G, T19H, T19I,T19K, T19L, T19M, T19N, T19P, T19Q, T19R, T19S, T19V, T19W, T19Y, G20*,G20A, G20D, G20E, G20F, G20H, G20I, G20K, G20L, G20M, G20N, G20P, G20Q,G20R, G20S, G20T, G20V, G20W, G20Y, S21*, S21A, S21D, S21E, S21F, S21G,S21H, S21I, S21K, S21L, S21M, S21N, S21P, S21Q, S21R, S21T, S21V, S21W,S21Y, G22*, G22A, G22D, G22E, G22F, G22H, G22I, G22K, G22L, G22M, G22N,G22P, G22Q, G22R, G22S, G22T, G22V, G22W, G22Y, D23*, D23A, D23E, D23F,D23G, D23H, D23I, D23K, D23L, D23M, D23N, D23P, D23Q, D23R, D23S, D23T,D23V, D23W, D23Y, S24*, S24A, S24D, S24E, S24F, S24G, S24H, S24I, S24K,S24L, S24M, S24N, S24P, S24Q, S24R, S24T, S24V, S24W, S24Y, Q25*, Q25A,Q25D, Q25E, Q25F, Q25G, Q25H, Q25I, Q25K, Q25L, Q25M, Q25N, Q25P, Q25R,Q25S, Q25T, Q25V, Q25W, Q25Y, S26*, S26A, S26D, S26E, S26F, S26G, S26H,S26I, S26K, S26L, S26M, S26N, S26P, S26Q, S26R, S26T, S26V, S26W, S26Y,D27*, D27A, D27E, D27F, D27G, D27H, D27I, D27K, D27L, D27M, D27N, D27P,D27Q, D27R, D27S, D27T, D27V, D27W, D27Y, P28*, P28A, P28D, P28E, P28F,P28G, P28H, P28I, P28K, P28L, P28M, P28N, P28Q, P28R, P28S, P28T, P28V,P28W, P28Y, I29*, I29A, I29D, I29E, I29F, I29G, I29H, I29K, I29L, I29M,I29N, I29P, I29Q, I29R, I29S, I29T, I29V, I29W, I29Y, K30*, K30A, K30D,K30E, K30F, K30G, K30H, K30I, K30L, K30M, K30N, K30P, K30Q, K30R, K30S,K30T, K30V, K30W, K30Y, A31*, A31D, A31E, A31F, A31G, A31H, A31I, A31K,A31L, A31M, A31N, A31P, A31Q, A31R, A31S, A31T, A31V, A31W, A31Y, D32*,D32A, D32E, D32F, D32G, D32H, D32I, D32K, D32L, D32M, D32N, D32P, D32Q,D32R, D32S, D32T, D32V, D32W, D32Y, L33*, L33A, L33D, L33E, L33F, L33G,L33H, L33I, L33K, L33M, L33N, L33P, L33Q, L33R, L33S, L33T, L33V, L33W,L33Y, E34*, E34A, E34D, E34F, E34G, E34H, E34I, E34K, E34L, E34M, E34N,E34P, E34Q, E34R, E34S, E34T, E34V, E34W, E34Y, V35*, V35A, V35D, V35E,V35F, V35G, V35H, V35I, V35K, V35L, V35M, V35N, V35P, V35Q, V35R, V35S,V35T, V35W, V35Y, K36*, K36A, K36D, K36E, K36F, K36G, K36H, K36I, K36L,K36M, K36N, K36P, K36Q, K36R, K36S, K36T, K36V, K36W, K36Y, G37*, G37A,G37D, G37E, G37F, G37H, G37I, G37K, G37L, G37M, G37N, G37P, G37Q, G37R,G37S, G37T, G37V, G37W, G37Y, Q38*, Q38A, Q38D, Q38E, Q38F, Q38G, Q38H,Q38I, Q38K, Q38L, Q38M, Q38N, Q38P, Q38R, Q38S, Q38T, Q38V, Q38W, Q38Y,S39*, S39A, S39D, S39E, S39F, S39G, S39H, S39I, S39K, S39L, S39M, S39N,S39P, S39Q, S39R, S39T, S39V, S39W, S39Y, A40*, A40D, A40E, A40F, A40G,A40H, A40I, A40K, A40L, A40M, A40N, A40P, A40Q, A40R, A40S, A40T, A40V,A40W, A40Y, L41*, L41A, L41D, L41E, L41F, L41G, L41H, L41I, L41K, L41M,L41N, L41P, L41Q, L41R, L41S, L41T, L41V, L41W, L41Y, P42*, P42A, P42D,P42E, P42F, P42G, P42H, P42I, P42K, P42L, P42M, P42N, P42Q, P42R, P42S,P42T, P42V, P42W, P42Y, F43*, F43A, F43D, F43E, F43G, F43H, F43I, F43K,F43L, F43M, F43N, F43P, F43Q, F43R, F43S, F43T, F43V, F43W, F43Y, D44*,D44A, D44E, D44F, D44G, D44H, D44I, D44K, D44L, D44M, D44N, D44P, D44Q,D44R, D44S, D44T, D44V, D44W, D44Y, V45*, V45A, V45D, V45E, V45F, V45G,V45H, V45I, V45K, V45L, V45M, V45N, V45P, V45Q, V45R, V45S, V45T, V45W,V45Y, D46*, D46A, D46E, D46F, D46G, D46H, D46I, D46K, D46L, D46M, D46N,D46P, D46Q, D46R, D46S, D46T, D46V, D46W, D46Y, C47A, C47D, C47E, C47F,C47G, C47H, C47I, C47K, C47L, C47M, C47N, C47P, C47Q, C47R, C47S, C47T,C47V, C47W, C47Y, W48*, W48A, W48D, W48E, W48F, W48G, W48H, W48I, W48K,W48L, W48M, W48N, W48P, W48Q, W48R, W48S, W48T, W48V, W48Y, A49*, A49D,A49E, A49F, A49G, A49H, A49I, A49K, A49L, A49M, A49N, A49P, A49Q, A49R,A49S, A49T, A49V, A49W, A49Y, I50*, I50A, I50D, I50E, I50F, I50G, I50H,I50K, I50L, I50M, I50N, I50P, I50Q, I50R, I50S, I50T, I50V, I50W, I50Y,L51*, L51A, L51D, L51E, L51F, L51G, L51H, L51I, L51K, L51M, L51N, L51P,L51Q, L51R, L51S, L51T, L51V, L51W, L51Y, K53*, K53A, K53D, K53E, K53F,K53G, K53H, K53I, K53L, K53M, K53N, K53P, K53Q, K53R, K53S, K53T, K53V,K53W, K53Y, G54*, G54A, G54D, G54E, G54F, G54H, G54I, G54K, G54L, G54M,G54N, G54P, G54Q, G54R, G54S, G54T, G54V, G54W, G54Y, A55*, A55D, A55E,A55F, A55G, A55H, A55I, A55K, A55L, A55M, A55N, A55P, A55Q, A55R, A55S,A55T, A55V, A55W, A55Y, P56*, P56A, P56D, P56E, P56F, P56G, P56H, P56I,P56K, P56L, P56M, P56N, P56Q, P56R, P56S, P56T, P56V, P56W, P56Y, N57*,N57A, N57D, N57E, N57F, N57G, N57H, N57I, N57K, N57L, N57M, N57P, N57Q,N57R, N57S, N57T, N57V, N57W, N57Y, V58*, V58A, V58D, V58E, V58F, V58G,V58H, V58I, V58K, V58L, V58M, V58N, V58P, V58Q, V58R, V58S, V58T, V58W,V58YL59*, L59A, L59D, L59E, L59F, L59G, L59H, L59I, L59K, L59M, L59N,L59P, L59Q, L59R, L59S, L59T, L59V, L59W, L59Y, Q60*, Q60A, Q60D, Q60E,Q60F, Q60G, Q60H, Q60I, Q60K, Q60L, Q60M, Q60N, Q60P, Q60R, Q60S, Q60T,Q60V, Q60W, Q60Y, R61*, R61A, R61D, R61E, R61F, R61G, R61H, R61I, R61K,R61L, R61M, R61N, R61P, R61Q, R61S, R61T, R61V, R61W, R61Y, V62*, V62A,V62D, V62E, V62F, V62G, V62H, V62I, V62K, V62L, V62M, V62N, V62P, V62Q,V62R, V62S, V62T, V62W, V62Y, N63*, N63A, N63D, N63E, N63F, N63G, N63H,N63I, N63K, N63L, N63M, N63P, N63Q, N63R, N63S, N63T, N63V, N63W, N63Y,E64*, E64A, E64D, E64F, E64G, E64H, E64I, E64K, E64L, E64M, E64N, E64P,E64Q, E64R, E64S, E64T, E64V, E64W, E64Y, K65*, K65A, K65D, K65E, K65F,K65G, K65H, K65I, K65L, K65M, K65N, K65P, K65Q, K65R, K65S, K65T, K65V,K65W, K65Y, T66*, T66A, T66D, T66E, T66F, T66G, T66H, T66I, T66K, T66L,T66M, T66N, T66P, T66Q, T66R, T66S, T66V, T66W, T66Y, K67*, K67A, K67D,K67E, K67F, K67G, K67H, K67I, K67L, K67M, K67N, K67P, K67Q, K67R, K67S,K67T, K67V, K67W, K67Y, N68*, N68A, N68D, N68E, N68F, N68G, N68H, N68I,N68K, N68L, N68M, N68P, N68Q, N68R, N68S, N68T, N68V, N68W, N68Y, S69*,S69A, S69D, S69E, S69F, S69G, S69H, S69I, S69K, S69L, S69M, S69N, S69P,S69Q, S69R, S69T, S69V, S69W, S69Y, N70*, N70A, N70D, N70E, N70F, N70G,N70H, N70I, N70K, N70L, N70M, N70P, N70Q, N70R, N70S, N70T, N70V, N70W,N70Y, R71*, R71A, R71D, R71E, R71F, R71G, R71H, R71I, R71K, R71L, R71M,R71N, R71P, R71Q, R71S, R71T, R71V, R71W, R71Y, D72*, D72A, D72E, D72F,D72G, D72H, D72I, D72K, D72L, D72M, D72N, D72P, D72Q, D72R, D72S, D72T,D72V, D72W, D72Y, R73*, R73A, R73D, R73E, R73F, R73G, R73H, R73I, R73K,R73L, R73M, R73N, R73P, R73Q, R73S, R73T, R73V, R73W, R73Y, S74*, S74A,S74D, S74E, S74F, S74G, S74H, S74I, S74K, S74L, S74M, S74N, S74P, S74Q,S74R, S74T, S74V, S74W, S74Y, G75*, G75A, G75D, G75E, G75F, G75H, G75I,G75K, G75L, G75M, G75N, G75P, G75Q, G75R, G75S, G75T, G75V, G75W, G75Y,A76*, A76D, A76E, A76F, A76G, A76H, A76I, A76K, A76L, A76M, A76N, A76P,A76Q, A76R, A76S, A76T, A76V, A76W, A76Y, N77*, N77A, N77D, N77E, N77F,N77G, N77H, N77I, N77K, N77L, N77M, N77P, N77Q, N77R, N77S, N77T, N77V,N77W, N77Y, K78*, K78A, K78D, K78E, K78F, K78G, K78H, K78I, K78L, K78M,K78N, K78P, K78Q, K78R, K78S, K78T, K78V, K78W, K78Y, G79*, G79A, G79D,G79E, G79F, G79H, G79I, G79K, G79L, G79M, G79N, G79P, G79Q, G79R, G79S,G79T, G79V, G79W, G79Y, P80*, P80A, P80D, P80E, P80F, P80G, P80H, P80I,P80K, P80L, P80M, P80N, P80Q, P80R, P80S, P80T, P80V, P80W, P80Y, F81*,F81A, F81D, F81E, F81G, F81H, F81I, F81K, F81L, F81M, F81N, F81P, F81Q,F81R, F81S, F81T, F81V, F81W, F81Y, K82*, K82A, K82D, K82E, K82F, K82G,K82H, K82I, K82L, K82M, K82N, K82P, K82Q, K82R, K82S, K82T, K82V, K82W,K82Y, D83*, D83A, D83E, D83F, D83G, D83H, D83I, D83K, D83L, D83M, D83N,D83P, D83Q, D83R, D83S, D83T, D83V, D83W, D83Y, P84*, P84A, P84D, P84E,P84F, P84G, P84H, P84I, P84K, P84L, P84M, P84N, P84Q, P84R, P84S, P84T,P84V, P84W, P84Y, Q85*, Q85A, Q85D, Q85E, Q85F, Q85G, Q85H, Q85I, Q85K,Q85L, Q85M, Q85N, Q85P, Q85R, Q85S, Q85T, Q85V, Q85W, Q85Y, K86*, K86A,K86D, K86E, K86F, K86G, K86H, K86I, K86L, K86M, K86N, K86P, K86Q, K86R,K86S, K86T, K86V, K86W, K86Y, W87*, W87A, W87D, W87E, W87F, W87G, W87H,W87I, W87K, W87L, W87M, W87N, W87P, W87Q, W87R, W87S, W87T, W87V, W87Y,G88*, G88A, G88D, G88E, G88F, G88H, G88I, G88K, G88L, G88M, G88N, G88P,G88Q, G88R, G88S, G88T, G88V, G88W, G88Y, I89*, I89A, I89D, I89E, I89F,I89G, I89H, I89K, I89L, I89M, I89N, I89P, I89Q, I89R, I89S, I89T, I89V,I89W, I89Y, K90*, K90A, K90D, K90E, K90F, K90G, K90H, K90I, K90L, K90M,K90N, K90P, K90Q, K90R, K90S, K90T, K90V, K90W, K90Y, A91*, A91D, A91E,A91F, A91G, A91H, A91I, A91K, A91L, A91M, A91N, A91P, A91Q, A91R, A91S,A91T, A91V, A91W, A91Y, L92*, L92A, L92D, L92E, L92F, L92G, L92H, L92I,L92K, L92M, L92N, L92P, L92Q, L92R, L92S, L92T, L92V, L92W, L92Y, P93*,P93A, P93D, P93E, P93F, P93G, P93H, P93I, P93K, P93L, P93M, P93N, P93Q,P93R, P93S, P93T, P93V, P93W, P93Y, P94*, P94A, P94D, P94E, P94F, P94G,P94H, P94I, P94K, P94L, P94M, P94N, P94Q, P94R, P94S, P94T, P94V, P94W,P94Y, K95*, K95A, K95D, K95E, K95F, K95G, K95H, K95I, K95L, K95M, K95N,K95P, K95Q, K95R, K95S, K95T, K95V, K95W, K95Y, N96*, N96A, N96D, N96E,N96F, N96G, N96H, N96I, N96K, N96L, N96M, N96P, N96Q, N96R, N96S, N96T,N96V, N96W, N96Y, P97*, P97A, P97D, P97E, P97F, P97G, P97H, P97I, P97K,P97L, P97M, P97N, P97Q, P97R, P97S, P97T, P97V, P97W, P97Y, S98*, S98A,S98D, S98E, S98F, S98G, S98H, S98I, S98K, S98L, S98M, S98N, S98P, S98Q,S98R, S98T, S98V, S98W, S98Y, W99*, W99A, W99D, W99E, W99F, W99G, W99H,W99I, W99K, W99L, W99M, W99N, W99P, W99Q, W99R, W99S, W99T, W99V, W99Y,S100*, S100A, S100D, S100E, S100F, S100G, S100H, S100I, S100K, S100L,S100M, S100N, S100P, S100Q, S100R, S100T, S100V, S100W, S100Y, A101*,A101D, A101E, A101F, A101G, A101H, A101I, A101K, A101L, A101M, A101N,A101P, A101Q, A101R, A101S, A101T, A101V, A101W, A101Y, Q102*, Q102A,Q102D, Q102E, Q102F, Q102G, Q102H, Q102I, Q102K, Q102L, Q102M, Q102N,Q102P, Q102R, Q102S, Q102T, Q102V, Q102W, Q102Y, D103*, D103A, D103E,D103F, D103G, D103H, D103I, D103K, D103L, D103M, D103N, D103P, D103Q,D103R, D103S, D103T, D103V, D103W, D103Y, F104*, F104A, F104D, F104E,F104G, F104H, F104I, F104K, F104L, F104M, F104N, F104P, F104Q, F104R,F104S, F104T, F104V, F104W, F104Y, K105*, K105A, K105D, K105E, K105F,K105G, K105H, K105I, K105L, K105M, K105N, K105P, K105Q, K105R, K105S,K105T, K105V, K105W, K105Y, S106*, S106A, S106D, S106E, S106F, S106G,S106H, S106I, S106K, S106L, S106M, S106N, S106P, S106Q, S106R, S106T,S106V, S106W, S106Y, P107*, P107A, P107D, P107E, P107F, P107G, P107H,P107I, P107K, P107L, P107M, P107N, P107Q, P107R, P107S, P107T, P107V,P107W, P107Y, E108*, E108A, E108D, E108F, E108G, E108H, E108I, E108K,E108L, E108M, E108N, E108P, E108Q, E108R, E108S, E108T, E108V, E108W,E108Y, E109*, E109A, E109D, E109F, E109G, E109H, E109I, E109K, E109L,E109M, E109N, E109P, E109Q, E109R, E109S, E109T, E109V, E109W, E109Y,Y110*, Y110A, Y110D, Y110E, Y110F, Y110G, Y110H, Y110I, Y110K, Y110L,Y110M, Y110N, Y110P, Y110Q, Y110R, Y110S, Y110T, Y110V, Y110W, A111*,A111D, A111E, A111F, A111G, A111H, A111I, A111K, A111L, A111M, A111N,A111P, A111Q, A111R, A111S, A111T, A111V, A111W, A111Y, F112*, F112A,F112D, F112E, F112G, F112H, F112I, F112K, F112L, F112M, F112N, F112P,F112Q, F112R, F112S, F112T, F112V, F112W, F112Y, A113*, A113D, A113E,A113F, A113G, A113H, A113I, A113K, A113L, A113M, A113N, A113P, A113Q,A113R, A113S, A113T, A113V, A113W, A113Y, S114*, S114A, S114D, S114E,S114F, S114G, S114H, S114I, S114K, S114L, S114M, S114N, S114P, S114Q,S114R, S114T, S114V, S114W, S114Y, S115*, S115A, S115D, S115E, S115F,S115G, S115H, S115I, S115K, S115L, S115M, S115N, S115P, S115Q, S115R,S115T, S115V, S115W, S115Y, L116*, L116A, L116D, L116E, L116F, L116G,L116H, L116I, L116K, L116M, L116N, L116P, L116Q, L116R, L116S, L116T,L116V, L116W, L116Y, Q117*, Q117A, Q117D, Q117E, Q117F, Q117G, Q117H,Q117I, Q117K, Q117L, Q117M, Q117N, Q117P, Q117R, Q117S, Q117T, Q117V,Q117W, Q117Y, G118*, G118A, G118D, G118E, G118F, G118H, G118I, G118K,G118L, G118M, G118N, G118P, G118Q, G118R, G118S, G118T, G118V, G118W,G118Y, G119*, G119A, G119D, G119E, G119F, G119H, G119I, G119K, G119L,G119M, G119N, G119P, G119Q, G119R, G119S, G119T, G119V, G119W, G119Y,T120*, T120A, T120D, T120E, T120F, T120G, T120H, T120I, T120K, T120L,T120M, T120N, T120P, T120Q, T120R, T120S, T120V, T120W, T120Y, N121*,N121A, N121D, N121E, N121F, N121G, N121H, N121I, N121K, N121L, N121M,N121P, N121Q, N121R, N121S, N121T, N121V, N121W, N121Y, A122*, A122D,A122E, A122F, A122G, A122H, A122I, A122K, A122L, A122M, A122N, A122P,A122Q, A122R, A122S, A122T, A122V, A122W, A122Y, I123*, I123A, I123D,I123E, I123F, I123G, I123H, I123K, I123L, I123M, I123N, I123P, I123Q,I123R, I123S, I123T, I123V, I123W, I123Y, L124*, L124A, L124D, L124E,L124F, L124G, L124H, L124I, L124K, L124M, L124N, L124P, L124Q, L124R,L124S, L124T, L124V, L124W, L124Y, A125*, A125D, A125E, A125F, A125G,A125H, A125I, A125K, A125L, A125M, A125N, A125P, A125Q, A125R, A125S,A125T, A125V, A125W, A125Y, P126*, P126A, P126D, P126E, P126F, P126G,P126H, P126I, P126K, P126L, P126M, P126N, P126Q, P126R, P126S, P126T,P126V, P126W, P126Y, V127*, V127A, V127D, V127E, V127F, V127G, V127H,V127I, V127K, V127L, V127M, V127N, V127P, V127Q, V127R, V127S, V127T,V127W, V127Y, N128*, N128A, N128D, N128E, N128F, N128G, N128H, N128I,N128K, N128L, N128M, N128P, N128Q, N128R, N128S, N128T, N128V, N128W,N128Y, L129*, L129A, L129D, L129E, L129F, L129G, L129H, L129I, L129K,L129M, L129N, L129P, L129Q, L129R, L129S, L129T, L129V, L129W, L129Y,A130*, A130D, A130E, A130F, A130G, A130H, A130I, A130K, A130L, A130M,A130N, A130P, A130Q, A130R, A130S, A130T, A130V, A130W, A130Y, S131*,S131A, S131D, S131E, S131F, S131G, S131H, S131I, S131K, S131L, S131M,S131N, S131P, S131Q, S131R, S131T, S131V, S131W, S131Y, Q132*, Q132A,Q132D, Q132E, Q132F, Q132G, Q132H, Q132I, Q132K, Q132L, Q132M, Q132N,Q132P, Q132R, Q132S, Q132T, Q132V, Q132W, Q132Y, N133*, N133A, N133D,N133E, N133F, N133G, N133H, N133I, N133K, N133L, N133M, N133P, N133Q,N133R, N133S, N133T, N133V, N133W, N133Y, S134*, S134A, S134D, S134E,S134F, S134G, S134H, S134I, S134K, S134L, S134M, S134N, S134P, S134Q,S134R, S134T, S134V, S134W, S134Y, Q135*, Q135A, Q135D, Q135E, Q135F,Q135G, Q135H, Q135I, Q135K, Q135L, Q135M, Q135N, Q135P, Q135R, Q135S,Q135T, Q135V, Q135W, Q135Y, G136*, G136A, G136D, G136E, G136F, G136H,G136I, G136K, G136L, G136M, G136N, G136P, G136Q, G136R, G136S, G136T,G136V, G136W, G136Y, G137*, G137A, G137D, G137E, G137F, G137H, G137I,G137K, G137L, G137M, G137N, G137P, G137Q, G137R, G137S, G137T, G137V,G137W, G137Y, V138*, V138A, V138D, V138E, V138F, V138G, V138H, V138I,V138K, V138L, V138M, V138N, V138P, V138Q, V138R, V138S, V138T, V138W,V138Y, L139*, L139A, L139D, L139E, L139F, L139G, L139H, L139I, L139K,L139M, L139N, L139P, L139Q, L139R, L139S, L139T, L139V, L139W, L139Y,N140*, N140A, N140D, N140E, N140F, N140G, N140H, N140I, N140K, N140L,N140M, N140P, N140Q, N140R, N140S, N140T, N140V, N140W, N140Y, G141*,G141A, G141D, G141E, G141F, G141H, G141I, G141K, G141L, G141M, G141N,G141P, G141Q, G141R, G141S, G141T, G141V, G141W, G141Y, F142*, F142A,F142D, F142E, F142G, F142H, F142I, F142K, F142L, F142M, F142N, F142P,F142Q, F142R, F142S, F142T, F142V, F142W, F142Y, Y143*, Y143A, Y143D,Y143E, Y143F, Y143G, Y143H, Y143I, Y143K, Y143L, Y143M, Y143N, Y143P,Y143Q, Y143R, Y143S, Y143T, Y143V, Y143W, S144*, S144A, S144D, S144E,S144F, S144G, S144H, S144I, S144K, S144L, S144M, S144N, S144P, S144Q,S144R, S144T, S144V, S144W, S144Y, A145*, A145D, A145E, A145F, A145G,A145H, A145I, A145K, A145L, A145M, A145N, A145P, A145Q, A145R, A145S,A145T, A145V, A145W, A145Y, N146*, N146A, N146D, N146E, N146F, N146G,N146H, N146I, N146K, N146L, N146M, N146P, N146Q, N146R, N146S, N146T,N146V, N146W, N146Y, K147*, K147A, K147D, K147E, K147F, K147G, K147H,K147I, K147L, K147M, K147N, K147P, K147Q, K147R, K147S, K147T, K147V,K147W, K147Y, V148*, V148A, V148D, V148E, V148F, V148G, V148H, V148I,V148K, V148L, V148M, V148N, V148P, V148Q, V148R, V148S, V148T, V148W,V148Y, A149*, A149D, A149E, A149F, A149G, A149H, A149I, A149K, A149L,A149M, A149N, A149P, A149Q, A149R, A149S, A149T, A149V, A149W, A149Y,Q150*, Q150A, Q150D, Q150E, Q150F, Q150G, Q150H, Q150I, Q150K, Q150L,Q150M, Q150N, Q150P, Q150R, Q150S, Q150T, Q150V, Q150W, Q150Y, F151*,F151A, F151D, F151E, F151G, F151H, F151I, F151K, F151L, F151M, F151N,F151P, F151Q, F151R, F151S, F151T, F151V, F151W, F151Y, D152*, D152A,D152E, D152F, D152G, D152H, D152I, D152K, D152L, D152M, D152N, D152P,D152Q, D152R, D152S, D152T, D152V, D152W, D152Y, P153*, P153A, P153D,P153E, P153F, P153G, P153H, P153I, P153K, P153L, P153M, P153N, P153Q,P153R, P153S, P153T, P153V, P153W, P153Y, S154*, S154A, S154D, S154E,S154F, S154G, S154H, S154I, S154K, S154L, S154M, S154N, S154P, S154Q,S154R, S154T, S154V, S154W, S154Y, K155*, K155A, K155D, K155E, K155F,K155G, K155H, K155I, K155L, K155M, K155N, K155P, K155Q, K155R, K155S,K155T, K155V, K155W, K155Y, P156*, P156A, P156D, P156E, P156F, P156G,P156H, P156I, P156K, P156L, P156M, P156N, P156Q, P156R, P156S, P156T,P156V, P156W, P156Y, Q157*, Q157A, Q157D, Q157E, Q157F, Q157G, Q157H,Q157I, Q157K, Q157L, Q157M, Q157N, Q157P, Q157R, Q157S, Q157T, Q157V,Q157W, Q157Y, Q158*, Q158A, Q158D, Q158E, Q158F, Q158G, Q158H, Q158I,Q158K, Q158L, Q158M, Q158N, Q158P, Q158R, Q158S, Q158T, Q158V, Q158W,Q158Y, T159*, T159A, T159D, T159E, T159F, T159G, T159H, T159I, T159K,T159L, T159M, T159N, T159P, T159Q, T159R, T159S, T159V, T159W, T159Y,K160*, K160A, K160D, K160E, K160F, K160G, K160H, K160I, K160L, K160M,K160N, K160P, K160Q, K160R, K160S, K160T, K160V, K160W, K160Y, G161*,G161A, G161D, G161E, G161F, G161H, G161I, G161K, G161L, G161M, G161N,G161P, G161Q, G161R, G161S, G161T, G161V, G161W, G161Y, T162A, T162D,T162E, T162F, T162G, T162H, T162I, T162K, T162L, T162M, T162N, T162P,T162Q, T162R, T162S, T162T, T162V, T162W, T162Y, W163*, W163A, W163D,W163E, W163F, W163G, W163H, W163I, W163K, W163L, W163M, W163N, W163P,W163Q, W163R, W163S, W163T, W163V, W163Y, F164*, F164A, F164D, F164E,F164G, F164H, F164I, F164K, F164L, F164M, F164N, F164P, F164Q, F164R,F164S, F164T, F164V, F164W, F164Y, Q165*, Q165A, Q165D, Q165E, Q165F,Q165G, Q165H, Q165I, Q165K, Q165L, Q165M, Q165N, Q165P, Q165R, Q165S,Q165T, Q165V, Q165W, Q165Y, I166*, I166A, I166D, I166E, I166F, I166G,I166H, I166K, I166L, I166M, I166N, I166P, I166Q, I166R, I166S, I166T,I166V, I166W, I166Y, T167*, T167A, T167D, T167E, T167F, T167G, T167H,T167I, T167K, T167L, T167M, T167N, T167P, T167Q, T167R, T167S, T167V,T167W, T167Y, K168*, K168A, K168D, K168E, K168F, K168G, K168H, K168I,K168L, K168M, K168N, K168P, K168Q, K168R, K168S, K168T, K168V, K168W,K168Y, F169*, F169A, F169D, F169E, F169G, F169H, F169I, F169K, F169L,F169M, F169N, F169P, F169Q, F169R, F169S, F169T, F169V, F169W, F169Y,T170*, T170A, T170D, T170E, T170F, T170G, T170H, T170I, T170K, T170L,T170M, T170N, T170P, T170Q, T170R, T170S, T170V, T170W, T170Y, G171*,G171A, G171D, G171E, G171F, G171H, G171I, G171K, G171L, G171M, G171N,G171P, G171Q, G171R, G171S, G171T, G171V, G171W, G171Y, A172*, A172D,A172E, A172F, A172G, A172H, A172I, A172K, A172L, A172M, A172N, A172P,A172Q, A172R, A172S, A172T, A172V, A172W, A172Y, A173*, A173D, A173E,A173F, A173G, A173H, A173I, A173K, A173L, A173M, A173N, A173P, A173Q,A173R, A173S, A173T, A173V, A173W, A173Y, G174*, G174A, G174D, G174E,G174F, G174H, G174I, G174K, G174L, G174M, G174N, G174P, G174Q, G174R,G174S, G174T, G174V, G174W, G174Y, P175*, P175A, P175D, P175E, P175F,P175G, P175H, P175I, P175K, P175L, P175M, P175N, P175Q, P175R, P175S,P175T, P175V, P175W, P175Y, Y176*, Y176A, Y176D, Y176E, Y176F, Y176G,Y176H, Y176I, Y176K, Y176L, Y176M, Y176N, Y176P, Y176Q, Y176R, Y176S,Y176T, Y176V, Y176W, K178*, K178A, K178D, K178E, K178F, K178G, K178H,K178I, K178L, K178M, K178N, K178P, K178Q, K178R, K178S, K178T, K178V,K178W, K178Y, A179*, A179D, A179E, A179F, A179G, A179H, A179I, A179K,A179L, A179M, A179N, A179P, A179Q, A179R, A179S, A179T, A179V, A179W,A179Y, L180*, L180A, L180D, L180E, L180F, L180G, L180H, L180I, L180K,L180M, L180N, L180P, L180Q, L180R, L180S, L180T, L180V, L180W, L180Y,G181*, G181A, G181D, G181E, G181F, G181H, G181I, G181K, G181L, G181M,G181N, G181P, G181Q, G181R, G181S, G181T, G181V, G181W, G181Y, S182*,S182A, S182D, S182E, S182F, S182G, S182H, S182I, S182K, S182L, S182M,S182N, S182P, S182Q, S182R, S182T, S182V, S182W, S182Y, N183*, N183A,N183D, N183E, N183F, N183G, N183H, N183I, N183K, N183L, N183M, N183P,N183Q, N183R, N183S, N183T, N183V, N183W, N183Y, D184*, D184A, D184E,D184F, D184G, D184H, D184I, D184K, D184L, D184M, D184N, D184P, D184Q,D184R, D184S, D184T, D184V, D184W, D184Y, K185*, K185A, K185D, K185E,K185F, K185G, K185H, K185I, K185L, K185M, K185N, K185P, K185Q, K185R,K185S, K185T, K185V, K185W, K185Y, S186*, S186A, S186D, S186E, S186F,S186G, S186H, S186I, S186K, S186L, S186M, S186N, S186P, S186Q, S186R,S186T, S186V, S186W, S186Y, V187*, V187A, V187D, V187E, V187F, V187G,V187H, V187I, V187K, V187L, V187M, V187N, V187P, V187Q, V187R, V187S,V187T, V187W, V187Y, D189*, D189A, D189E, D189F, D189G, D189H, D189I,D189K, D189L, D189M, D189N, D189P, D189Q, D189R, D189S, D189T, D189V,D189W, D189Y, K190*, K190A, K190D, K190E, K190F, K190G, K190H, K190I,K190L, K190M, K190N, K190P, K190Q, K190R, K190S, K190T, K190V, K190W,K190Y, N191*, N191A, N191D, N191E, N191F, N191G, N191H, N191I, N191K,N191L, N191M, N191P, N191Q, N191R, N191S, N191T, N191V, N191W, N191Y,K192*, K192A, K192D, K192E, K192F, K192G, K192H, K192I, K192L, K192M,K192N, K192P, K192Q, K192R, K192S, K192T, K192V, K192W, K192Y, N193*,N193A, N193D, N193E, N193F, N193G, N193H, N193I, N193K, N193L, N193M,N193P, N193Q, N193R, N193S, N193T, N193V, N193W, N193Y, I194*, I194A,I194D, I194E, I194F, I194G, I194H, I194K, I194L, I194M, I194N, I194P,I194Q, I194R, I194S, I194T, I194V, I194W, I194Y, A195*, A195D, A195E,A195F, A195G, A195H, A195I, A195K, A195L, A195M, A195N, A195P, A195Q,A195R, A195S, A195T, A195V, A195W, A195Y, G196*, G196A, G196D, G196E,G196F, G196H, G196I, G196K, G196L, G196M, G196N, G196P, G196Q, G196R,G196S, G196T, G196V, G196W, G196Y, D197*, D197A, D197E, D197F, D197G,D197H, D197I, D197K, D197L, D197M, D197N, D197P, D197Q, D197R, D197S,D197T, D197V, D197W, D197Y, W198*, W198A, W198D, W198E, W198F, W198G,W198H, W198I, W198K, W198L, W198M, W198N, W198P, W198Q, W198R, W198S,W198T, W198V, W198Y, G199*, G199A, G199D, G199E, G199F, G199H, G199I,G199K, G199L, G199M, G199N, G199P, G199Q, G199R, G199S, G199T, G199V,G199W, G199Y, F200*, F200A, F200D, F200E, F200G, F200H, F200I, F200K,F200L, F200M, F200N, F200P, F200Q, F200R, F200S, F200T, F200V, F200W,F200Y, D201*, D201A, D201E, D201F, D201G, D201H, D201I, D201K, D201L,D201M, D201N, D201P, D201Q, D201R, D201S, D201T, D201V, D201W, D201Y,P202*, P202A, P202D, P202E, P202F, P202G, P202H, P202I, P202K, P202L,P202M, P202N, P202Q, P202R, P202S, P202T, P202V, P202W, P202Y, A203*,A203D, A203E, A203F, A203G, A203H, A203I, A203K, A203L, A203M, A203N,A203P, A203Q, A203R, A203S, A203T, A203V, A203W, A203Y, K204*, K204A,K204D, K204E, K204F, K204G, K204H, K204I, K204L, K204M, K204N, K204P,K204Q, K204R, K204S, K204T, K204V, K204W, K204Y, W205*, W205A, W205D,W205E, W205F, W205G, W205H, W205I, W205K, W205L, W205M, W205N, W205P,W205Q, W205R, W205S, W205T, W205V, W205Y, A206*, A206D, A206E, A206F,A206G, A206H, A206I, A206K, A206L, A206M, A206N, A206P, A206Q, A206R,A206S, A206T, A206V, A206W, A206Y, Y207*, Y207A, Y207D, Y207E, Y207F,Y207G, Y207H, Y207I, Y207K, Y207L, Y207M, Y207N, Y207P, Y207Q, Y207R,Y207S, Y207T, Y207V, Y207W, Q208*, Q208A, Q208D, Q208E, Q208F, Q208G,Q208H, Q208I, Q208K, Q208L, Q208M, Q208N, Q208P, Q208R, Q208S, Q208T,Q208V, Q208W, Q208Y, Y209*, Y209A, Y209D, Y209E, Y209F, Y209G, Y209H,Y209I, Y209K, Y209L, Y209M, Y209N, Y209P, Y209Q, Y209R, Y209S, Y209T,Y209V, Y209W, D210*, D210A, D210E, D210F, D210G, D210H, D210I, D210K,D210L, D210M, D210N, D210P, D210Q, D210R, D210S, D210T, D210V, D210W,D210Y, E211*, E211A, E211D, E211F, E211G, E211H, E211I, E211K, E211L,E211M, E211N, E211P, E211Q, E211R, E211S, E211T, E211V, E211W, E211Y,K212*, K212A, K212D, K212E, K212F, K212G, K212H, K212I, K212L, K212M,K212N, K212P, K212Q, K212R, K212S, K212T, K212V, K212W, K212Y, N213*,N213A, N213D, N213E, N213F, N213G, N213H, N213I, N213K, N213L, N213M,N213P, N213Q, N213R, N213S, N213T, N213V, N213W, N213Y, N214*, N214A,N214D, N214E, N214F, N214G, N214H, N214I, N214K, N214L, N214M, N214P,N214Q, N214R, N214S, N214T, N214V, N214W, N214Y, K215*, K215A, K215D,K215E, K215F, K215G, K215H, K215I, K215L, K215M, K215N, K215P, K215Q,K215R, K215S, K215T, K215V, K215W, K215Y, F216*, F216A, F216D, F216E,F216G, F216H, F216I, F216K, F216L, F216M, F216N, F216P, F216Q, F216R,F216S, F216T, F216V, F216W, F216Y, N217*, N217A, N217D, N217E, N217F,N217G, N217H, N217I, N217K, N217L, N217M, N217P, N217Q, N217R, N217S,N217T, N217V, N217W, N217Y, Y218*, Y218A, Y218D, Y218E, Y218F, Y218G,Y218H, Y218I, Y218K, Y218L, Y218M, Y218N, Y218P, Y218Q, Y218R, Y218S,Y218T, Y218V, Y218W, V219*, V219A, V219D, V219E, V219F, V219G, V219H,V219I, V219K, V219L, V219M, V219N, V219P, V219Q, V219R, V219S, V219T,V219W, V219Y, G220*, G220A, G220D, G220E, G220F, G220H, G220I, G220K,G220L, G220M, G220N, K221F, K221G, K221H, K221I, K221L, K221M, K221N,K221P, K221Q, K221R, K221S, K221T, K221V, K221W and K221Y, wherein eachposition corresponds to the position of the polypeptide of SEQ ID NO 1,and wherein the variant have improved activity as compared to thereference DNase e.g. SEQ ID NO 1, when measured in the activity Assay Ias described in Assays and Detergents below.

One embodiment of the invention relates to DNase variants having atleast 60% identity to SEQ ID NO: 1, having DNase activity and comprisingone or more substitutions selected from the group consisting of V1A,V1D, V1E, V1F, V1G, V1H, V1I, V1K, V1L, V1M, V1N, V1P, V1Q, V1R, V1S,V1T, V1W, V1Y, P2A, P2D, P2E, P2F, P2G, P2H, P2I, P2K, P2L, P2M, P2N,P2Q, P2R, P2S, P2T, P2V, P2W, P2Y, V3A, V3C, V3D, V3E, V3F, V3G, V3H,V3I, V3K, V3L, V3M, V3N, V3P, V3R, V3S, V3T, V3W, V3Y, N4A, N4D, N4E,N4F, N4G, N4H, N4I, N4K, N4L, N4M, N4P, N4Q, N4R, N4S, N4T, N4V, N4W,N4Y, P5A, P5D, P5E, P5F, P5G, P5H, P5I, P5K, P5L, P5M, P5N, P5Q, P5R,P5S, P5T, P5V, P5W, P5Y, E6A, E6D, E6F, E6G, E6H, E6I, E6K, E6L, E6M,E6N, E6P, E6Q, E6R, E6S, E6T, E6V, E6W, E6Y, P7A, P7D, P7E, P7F, P7G,P7H, P7I, P7K, P7L, P7M, P7N, P7Q, P7R, P7S, P7T, P7V, P7W, P7Y, D8A,D8E, D8F, D8G, D8H, D8I, D8K, D8L, D8M, D8N, D8P, D8Q, D8R, D8S, D8T,D8V, D8W, D8Y, A9D, A9E, A9F, A9G, A9H, A9I, A9K, A9L, A9M, A9N, A9P,A9Q, A9R, A9S, A9T, A9V, A9W, A9Y, T10A, T10D, T10E, T10F, T10G, T10H,T10I, T10K, T10L, T10M, T10N, T10P, T10Q, T10R, T10S, T10V, T10W, T10Y,S11A, S11D, S11E, S11F, S11G, S11H, S11I, S11K, S11L, S11M, S11N, S11P,S11Q, S11R, S11T, S11V, S11W, S11Y, V12A, V12D, V12E, V12F, V12G, V12H,V12I, V12K, V12L, V12M, V12N, V12P, V12Q, V12R, V12S, V12T, V12W, V12Y,E13A, E13D, E13F, E13G, E13H, E13I, E13K, E13L, E13M, E13N, E13P, E13Q,E13R, E13S, E13T, E13V, E13W, E13Y, N14A, N14D, N14E, N14F, N14G, N14H,N14I, N14K, N14L, N14M, N14P, N14Q, N14R, N14S, N14T, N14V, N14W, N14Y,V15A, V15D, V15E, V15F, V15G, V15H, V15I, V15K, V15L, V15M, V15N, V15P,V15Q, V15R, V15S, V15T, V15W, V15Y, A16D, A16E, A16F, A16G, A16H, A16I,A16K, A16L, A16M, A16N, A16P, A16Q, A16R, A16S, A16T, A16V, A16W, A16Y,L17A, L17D, L17E, L17F, L17G, L17H, L17I, L17K, L17M, L17N, L17P, L17Q,L17R, L17S, L17T, L17V, L17W, L17Y, K18A, K18D, K18E, K18F, K18G, K18H,K18I, K18L, K18M, K18N, K18P, K18Q, K18R, K18S, K18T, K18V, K18W, K18Y,T19A, T19D, T19E, T19F, T19G, T19H, T19I, T19K, T19L, T19M, T19N, T19P,T19Q, T19R, T19S, T19V, T19W, T19Y, G20A, G20D, G20E, G20F, G20H, G20I,G20K, G20L, G20M, G20N, G20P, G20Q, G20R, G20S, G20T, G20V, G20W, G20Y,S21A, S21D, S21E, S21F, S21G, S21H, S21I, S21K, S21L, S21M, S21N, S21P,S21Q, S21R, S21T, S21V, S21W, S21Y, G22A, G22D, G22E, G22F, G22H, G22I,G22K, G22L, G22M, G22N, G22P, G22Q, G22R, G22S, G22T, G22V, G22W, G22Y,D23A, D23E, D23F, D23G, D23H, D23I, D23K, D23L, D23M, D23N, D23P, D23Q,D23R, D23S, D23T, D23V, D23W, D23Y, S24A, S24D, S24E, S24F, S24G, S24H,S24I, S24K, S24L, S24M, S24N, S24P, S24Q, S24R, S24T, S24V, S24W, S24Y,Q25A, Q25D, Q25E, Q25F, Q25G, Q25H, Q25I, Q25K, Q25L, Q25M, Q25N, Q25P,Q25R, Q25S, Q25T, Q25V, Q25W, Q25Y, S26A, S26D, S26E, S26F, S26G, S26H,S26I, S26K, S26L, S26M, S26N, S26P, S26Q, S26R, S26T, S26V, S26W, S26Y,D27A, D27E, D27F, D27G, D27H, D27I, D27K, D27L, D27M, D27N, D27P, D27Q,D27R, D27S, D27T, D27V, D27W, D27Y, P28A, P28D, P28E, P28F, P28G, P28H,P28I, P28K, P28L, P28M, P28N, P28Q, P28R, P28S, P28T, P28V, P28W, P28Y,I29A, I29D, I29E, I29F, I29G, I29H, I29K, I29L, I29M, I29N, I29P, I29Q,I29R, I29S, I29T, I29V, I29W, I29Y, K30A, K30D, K30E, K30F, K30G, K30H,K30I, K30L, K30M, K30N, K30P, K30Q, K30R, K30S, K30T, K30V, K30W, K30Y,A31D, A31E, A31F, A31G, A31H, A31I, A31K, A31L, A31M, A31N, A31P, A31Q,A31R, A31S, A31T, A31V, A31W, A31Y, D32A, D32E, D32F, D32G, D32H, D32I,D32K, D32L, D32M, D32N, D32P, D32Q, D32R, D32S, D32T, D32V, D32W, D32Y,L33A, L33D, L33E, L33F, L33G, L33H, L33I, L33K, L33M, L33N, L33P, L33Q,L33R, L33S, L33T, L33V, L33W, L33Y, E34A, E34D, E34F, E34G, E34H, E34I,E34K, E34L, E34M, E34N, E34P, E34Q, E34R, E34S, E34T, E34V, E34W, E34Y,V35A, V35D, V35E, V35F, V35G, V35H, V35I, V35K, V35L, V35M, V35N, V35P,V35Q, V35R, V35S, V35T, V35W, V35Y, K36A, K36D, K36E, K36F, K36G, K36H,K36I, K36L, K36M, K36N, K36P, K36Q, K36R, K36S, K36T, K36V, K36W, K36Y,G37A, G37D, G37E, G37F, G37H, G37I, G37K, G37L, G37M, G37N, G37P, G37Q,G37R, G37S, G37T, G37V, G37W, G37Y, Q38A, Q38D, Q38E, Q38F, Q38G, Q38H,Q38I, Q38K, Q38L, Q38M, Q38N, Q38P, Q38R, Q38S, Q38T, Q38V, Q38W, Q38Y,S39A, S39D, S39E, S39F, S39G, S39H, S39I, S39K, S39L, S39M, S39N, S39P,S39Q, S39R, S39T, S39V, S39W, S39Y, A40D, A40E, A40F, A40G, A40H, A40I,A40K, A40L, A40M, A40N, A40P, A40Q, A40R, A40S, A40T, A40V, A40W, A40Y,L41A, L41D, L41E, L41F, L41G, L41H, L41I, L41K, L41M, L41N, L41P, L41Q,L41R, L41S, L41T, L41V, L41W, L41Y, P42A, P42D, P42E, P42F, P42G, P42H,P42I, P42K, P42L, P42M, P42N, P42Q, P42R, P42S, P42T, P42V, P42W, P42Y,F43A, F43D, F43E, F43G, F43H, F43I, F43K, F43L, F43M, F43N, F43P, F43Q,F43R, F43S, F43T, F43V, F43W, F43Y, D44A, D44E, D44F, D44G, D44H, D44I,D44K, D44L, D44M, D44N, D44P, D44Q, D44R, D44S, D44T, D44V, D44W, D44Y,V45A, V45D, V45E, V45F, V45G, V45H, V45I, V45K, V45L, V45M, V45N, V45P,V45Q, V45R, V45S, V45T, V45W, V45Y, D46A, D46E, D46F, D46G, D46H, D46I,D46K, D46L, D46M, D46N, D46P, D46Q, D46R, D46S, D46T, D46V, D46W, D46Y,C47A, C47D, C47E, C47F, C47G, C47H, C47I, C47K, C47L, C47M, C47N, C47P,C47Q, C47R, C47S, C47T, C47V, C47W, C47Y, W48A, W48D, W48E, W48F, W48G,W48H, W48I, W48K, W48L, W48M, W48N, W48P, W48Q, W48R, W48S, W48T, W48V,W48Y, A49D, A49E, A49F, A49G, A49H, A49I, A49K, A49L, A49M, A49N, A49P,A49Q, A49R, A49S, A49T, A49V, A49W, A49Y, I50A, I50D, I50E, I50F, I50G,I50H, I50K, I50L, I50M, I50N, I50P, I50Q, I50R, I50S, I50T, I50V, I50W,I50Y, L51A, L51D, L51E, L51F, L51G, L51H, L51I, L51K, L51M, L51N, L51P,L51Q, L51R, L51S, L51T, L51V, L51W, L51Y, K53A, K53D, K53E, K53F, K53G,K53H, K53I, K53L, K53M, K53N, K53P, K53Q, K53R, K53S, K53T, K53V, K53W,K53Y, G54A, G54D, G54E, G54F, G54H, G54I, G54K, G54L, G54M, G54N, G54P,G54Q, G54R, G54S, G54T, G54V, G54W, G54Y, A55D, A55E, A55F, A55G, A55H,A55I, A55K, A55L, A55M, A55N, A55P, A55Q, A55R, A55S, A55T, A55V, A55W,A55Y, P56A, P56D, P56E, P56F, P56G, P56H, P56I, P56K, P56L, P56M, P56N,P56Q, P56R, P56S, P56T, P56V, P56W, P56Y, N57A, N57D, N57E, N57F, N57G,N57H, N57I, N57K, N57L, N57M, N57P, N57Q, N57R, N57S, N57T, N57V, N57W,N57Y, V58A, V58D, V58E, V58F, V58G, V58H, V58I, V58K, V58L, V58M, V58N,V58P, V58Q, V58R, V58S, V58T, V58W, V58Y, L59A, L59D, L59E, L59F, L59G,L59H, L59I, L59K, L59M, L59N, L59P, L59Q, L59R, L59S, L59T, L59V, L59W,L59Y, Q60A, Q60D, Q60E, Q60F, Q60G, Q60H, Q60I, Q60K, Q60L, Q60M, Q60N,Q60P, Q60R, Q60S, Q60T, Q60V, Q60W, Q60Y, R61A, R61D, R61E, R61F, R61G,R61H, R61I, R61K, R61L, R61M, R61N, R61P, R61Q, R61S, R61T, R61V, R61W,R61Y, V62A, V62D, V62E, V62F, V62G, V62H, V62I, V62K, V62L, V62M, V62N,V62P, V62Q, V62R, V62S, V62T, V62W, V62Y, N63A, N63D, N63E, N63F, N63G,N63H, N63I, N63K, N63L, N63M, N63P, N63Q, N63R, N63S, N63T, N63V, N63W,N63Y, E64A, E64D, E64F, E64G, E64H, E64I, E64K, E64L, E64M, E64N, E64P,E64Q, E64R, E64S, E64T, E64V, E64W, E64Y, K65A, K65D, K65E, K65F, K65G,K65H, K65I, K65L, K65M, K65N, K65P, K65Q, K65R, K65S, K65T, K65V, K65W,K65Y, T66A, T66D, T66E, T66F, T66G, T66H, T66I, T66K, T66L, T66M, T66N,T66P, T66Q, T66R, T66S, T66V, T66W, T66Y, K67A, K67D, K67E, K67F, K67G,K67H, K67I, K67L, K67M, K67N, K67P, K67Q, K67R, K67S, K67T, K67V, K67W,K67Y, N68A, N68D, N68E, N68F, N68G, N68H, N68I, N68K, N68L, N68M, N68P,N68Q, N68R, N68S, N68T, N68V, N68W, N68Y, S69A, S69D, S69E, S69F, S69G,S69H, S69I, S69K, S69L, S69M, S69N, S69P, S69Q, S69R, S69T, S69V, S69W,S69Y, N70A, N70D, N70E, N70F, N70G, N70H, N70I, N70K, N70L, N70M, N70P,N70Q, N70R, N70S, N70T, N70V, N70W, N70Y, R71A, R71D, R71E, R71F, R71G,R71H, R71I, R71K, R71L, R71M, R71N, R71P, R71Q, R71S, R71T, R71V, R71W,R71Y, D72A, D72E, D72F, D72G, D72H, D72I, D72K, D72L, D72M, D72N, D72P,D72Q, D72R, D72S, D72T, D72V, D72W, D72Y, R73A, R73D, R73E, R73F, R73G,R73H, R73I, R73K, R73L, R73M, R73N, R73P, R73Q, R73S, R73T, R73V, R73W,R73Y, S74A, S74D, S74E, S74F, S74G, S74H, S74I, S74K, S74L, S74M, S74N,S74P, S74Q, S74R, S74T, S74V, S74W, S74Y, G75A, G75D, G75E, G75F, G75H,G75I, G75K, G75L, G75M, G75N, G75P, G75Q, G75R, G75S, G75T, G75V, G75W,G75Y, A76D, A76E, A76F, A76G, A76H, A76I, A76K, A76L, A76M, A76N, A76P,A76Q, A76R, A76S, A76T, A76V, A76W, A76Y, N77A, N77D, N77E, N77F, N77G,N77H, N77I, N77K, N77L, N77M, N77P, N77Q, N77R, N77S, N77T, N77V, N77W,N77Y, K78A, K78D, K78E, K78F, K78G, K78H, K78I, K78L, K78M, K78N, K78P,K78Q, K78R, K78S, K78T, K78V, K78W, K78Y, G79A, G79D, G79E, G79F, G79H,G79I, G79K, G79L, G79M, G79N, G79P, G79Q, G79R, P80M, P80N, P80Q, P80R,P80S, P80T, P80V, P80W, P80Y, F81A, F81D, F81E, F81G, F81H, F81I, F81K,F81L, F81M, F81N, F81P, F81Q, F81R, F81S, F81T, F81V, F81W, F81Y, K82A,K82D, K82E, K82F, K82G, K82H, K82I, K82L, K82M, K82N, K82P, K82Q, K82R,K82S, K82T, K82V, K82W, K82Y, D83A, D83E, D83F, D83G, D83H, D83I, D83K,D83L, D83M, D83N, D83P, D83Q, D83R, D83S, D83T, D83V, D83W, D83Y, P84A,P84D, P84E, P84F, P84G, P84H, P84I, P84K, P84L, P84M, P84N, P84Q, P84R,P84S, P84T, P84V, P84W, P84Y, Q85A, Q85D, Q85E, Q85F, Q85G, Q85H, Q85I,Q85K, Q85L, Q85M, Q85N, Q85P, Q85R, Q85S, Q85T, Q85V, Q85W, Q85Y, K86A,K86D, K86E, K86F, K86G, K86H, K86I, K86L, K86M, K86N, K86P, K86Q, K86R,K86S, K86T, K86V, K86W, K86Y, W87A, W87D, W87E, W87F, W87G, W87H, W87I,W87K, W87L, W87M, W87N, W87P, W87Q, W87R, W87S, W87T, W87V, W87Y, G88A,G88D, G88E, G88F, G88H, G88I, G88K, G88L, G88M, G88N, G88P, G88Q, G88R,G88S, G88T, G88V, G88W, G88Y, I89A, I89D, I89E, I89F, I89G, I89H, I89K,I89L, I89M, I89N, I89P, I89Q, I89R, I89S, I89T, I89V, I89W, I89Y, K90A,K90D, K90E, K90F, K90G, K90H, K90I, K90L, K90M, K90N, K90P, K90Q, K90R,K90S, K90T, K90V, K90W, K90Y, A91D, A91E, A91F, A91G, A91H, A91I, A91K,A91L, A91M, A91N, A91P, A91Q, A91R, A91S, A91T, A91V, A91W, A91Y, L92A,L92D, L92E, L92F, L92G, L92H, L92I, L92K, L92M, L92N, L92P, L92Q, L92R,L92S, L92T, L92V, L92W, L92Y, P93A, P93D, P93E, P93F, P93G, P93H, P93I,P93K, P93L, P93M, P93N, P93Q, P93R, P93S, P93T, P93V, P93W, P93Y, P94A,P94D, P94E, P94F, P94G, P94H, P94I, P94K, P94L, P94M, P94N, P94Q, P94R,P94S, P94T, P94V, P94W, P94Y, K95A, K95D, K95E, K95F, K95G, K95H, K95I,K95L, K95M, K95N, K95P, K95Q, K95R, K95S, K95T, K95V, K95W, K95Y, N96A,N96D, N96E, N96F, N96G, N96H, N96I, N96K, N96L, N96M, N96P, N96Q, N96R,N96S, N96T, N96V, N96W, N96Y, P97A, P97D, P97E, P97F, P97G, P97H, P97I,P97K, P97L, P97M, P97N, P97Q, P97R, P97S, P97T, P97V, P97W, P97Y, S98A,S98D, S98E, S98F, S98G, S98H, S98I, S98K, S98L, S98M, S98N, S98P, S98Q,S98R, S98T, S98V, S98W, S98Y, W99A, W99D, W99E, W99F, W99G, W99H, W99I,W99K, W99L, W99M, W99N, W99P, W99Q, W99R, W99S, W99T, W99V, W99Y, S100A,S100D, S100E, S100F, S100G, S100H, S100I, S100K, S100L, S100M, S100N,S100P, S100Q, S100R, S100T, S100V, S100W, S100Y, A101D, A101E, A101F,A101G, A101H, A101I, A101K, A101L, A101M, A101N, A101P, A101Q, A101R,A101S, A101T, A101V, A101W, A101Y, Q102A, Q102D, Q102E, Q102F, Q102G,Q102H, Q102I, Q102K, Q102L, Q102M, Q102N, Q102P, Q102R, Q102S, Q102T,Q102V, Q102W, Q102Y, D103A, D103E, D103F, D103G, D103H, D103I, D103K,D103L, D103M, D103N, D103P, D103Q, D103R, D103S, D103T, D103V, D103W,D103Y, F104A, F104D, F104E, F104G, F104H, F104I, F104K, F104L, F104M,F104N, F104P, F104Q, F104R, F104S, F104T, F104V, F104W, F104Y, K105A,K105D, K105E, K105F, K105G, K105H, K105I, K105L, K105M, K105N, K105P,K105Q, K105R, K105S, K105T, K105V, K105W, K105Y, S106A, S106D, S106E,S106F, S106G, S106H, S106I, S106K, S106L, S106M, S106N, S106P, S106Q,S106R, S106T, S106V, S106W, S106Y, P107A, P107D, P107E, P107F, P107G,P107H, P107I, P107K, P107L, P107M, P107N, P107Q, P107R, P107S, P107T,P107V, P107W, P107Y, E108A, E108D, E108F, E108G, E108H, E108I, E108K,E108L, E108M, E108N, E108P, E108Q, E108R, E108S, E108T, E108V, E108W,E108Y, E109A, E109D, E109F, E109G, E109H, E109I, E109K, E109L, E109M,E109N, E109P, E109Q, E109R, E109S, E109T, E109V, E109W, E109Y, Y110A,Y110D, Y110E, Y110F, Y110G, Y110H, Y110I, Y110K, Y110L, Y110M, Y110N,Y110P, Y110Q, Y110R, Y110S, Y110T, Y110V, Y110W, A111D, A111E, A111F,A111G, A111H, A111I, A111K, A111L, A111M, A111N, A111P, A111Q, A111R,A111S, A111T, A111V, A111W, A111Y, F112A, F112D, F112E, F112G, F112H, PPF112I, F112K, F112L, F112M, F112N, F112P, F112Q, F112R, F112S, F112T,F112V, F112W, F112Y, A113D, A113E, A113F, A113G, A113H, A113I, A113K,A113L, A113M, A113N, A113P, A113Q, A113R, A113S, A113T, A113V, A113W,A113Y, S114A, S114D, S114E, S114F, S114G, S114H, S114I, S114K, S114L,S114M, S114N, S114P, S114Q, S114R, S114T, S114V, S114W, S114Y, S115A,S115D, S115E, S115F, S115G, S115H, S115I, S115K, S115L, S115M, S115N,S115P, S115Q, S115R, S115T, S115V, S115W, S115Y, L116A, L116D, L116E,L116F, L116G, L116H, L116I, L116K, L116M, L116N, L116P, L116Q, L116R,L116S, L116T, L116V, L116W, L116Y, Q117A, Q117D, Q117E, Q117F, Q117G,Q117H, Q117I, Q117K, Q117L, Q117M, Q117N, Q117P, Q117R, Q117S, Q117T,Q117V, Q117W, Q117Y, G118A, G118D, G118E, G118F, G118H, G118I, G118K,G118L, G118M, G118N, G118P, G118Q, G118R, G118S, G118T, G118V, G118W,G118Y, G119A, G119D, G119E, G119F, G119H, G119I, G119K, G119L, G119M,G119N, G119P, G119Q, G119R, G119S, G119T, G119V, G119W, G119Y, T120A,T120D, T120E, T120F, T120G, T120H, T120I, T120K, T120L, T120M, T120N,T120P, T120Q, T120R, T120S, T120V, T120W, T120Y, N121A, N121D, N121E,N121F, N121G, N121H, N121I, N121K, N121L, N121M, N121P, N121Q, N121R,N121S, N121T, N121V, N121W, N121Y, A122D, A122E, A122F, A122G, A122H,A122I, A122K, A122L, A122M, A122N, A122P, A122Q, A122R, A122S, A122T,A122V, A122W, A122Y, I123A, I123D, I123E, I123F, I123G, I123H, I123K,I123L, I123M, I123N, I123P, I123Q, I123R, I123S, I123T, I123V, I123W,I123Y, L124A, L124D, L124E, L124F, L124G, L124H, L124I, L124K, L124M,L124N, L124P, L124Q, L124R, L124S, L124T, L124V, L124W, L124Y, A125D,A125E, A125F, A125G, A125H, A125I, A125K, A125L, A125M, A125N, A125P,A125Q, A125R, A125S, A125T, A125V, A125W, A125Y, P126A, P126D, P126E,P126F, P126G, P126H, P126I, P126K, P126L, P126M, P126N, P126Q, P126R,P126S, P126T, P126V, P126W, P126Y, V127A, V127D, V127E, V127F, V127G,V127H, V127I, V127K, V127L, V127M, V127N, V127P, V127Q, V127R, V127S,V127T, V127W, V127Y, N128A, N128D, N128E, N128F, N128G, N128H, N128I,N128K, N128L, N128M, N128P, N128Q, N128R, N128S, N128T, N128V, N128W,N128Y, L129A, L129D, L129E, L129F, L129G, L129H, L129I, L129K, L129M,L129N, L129P, L129Q, L129R, L129S, L129T, L129V, L129W, L129Y, A130D,A130E, A130F, A130G, A130H, A130I, A130K, A130L, A130M, A130N, A130P,A130Q, A130R, A130S, A130T, A130V, A130W, A130Y, S131A, S131D, S131E,S131F, S131G, S131H, S131I, S131K, S131L, S131M, S131N, S131P, S131Q,S131R, S131T, S131V, S131W, S131Y, Q132A, Q132D, Q132E, Q132F, Q132G,Q132H, Q132I, Q132K, Q132L, Q132M, Q132N, Q132P, Q132R, Q132S, Q132T,Q132V, Q132W, Q132Y, N133A, N133D, N133E, N133F, N133G, N133H, N133I,N133K, N133L, N133M, N133P, N133Q, N133R, N133S, N133T, N133V, N133W,N133Y, S134A, S134D, S134E, S134F, S134G, S134H, S134I, S134K, S134L,S134M, S134N, S134P, S134Q, S134R, S134T, S134V, S134W, S134Y, Q135A,Q135D, Q135E, Q135F, Q135G, Q135H, Q135I, Q135K, Q135L, Q135M, Q135N,Q135P, Q135R, Q135S, Q135T, Q135V, Q135W, Q135Y, G136A, G136D, G136E,G136F, G136H, G136I, G136K, G136L, G136M, G136N, G136P, G136Q, G136R,G136S, G136T, G136V, G136W, G136Y, G137A, G137D, G137E, G137F, G137H,G137I, G137K, G137L, G137M, G137N, G137P, G137Q, G137R, G137S, G137T,G137V, G137W, G137Y, V138A, V138D, V138E, V138F, V138G, V138H, V138I,V138K, V138L, V138M, V138N, V138P, V138Q, V138R, V138S, V138T, V138W,V138Y, L139A, L139D, L139E, L139F, L139G, L139H, L139I, L139K, L139M,L139N, L139P, L139Q, L139R, L139S, L139T, L139V, L139W, L139Y, N140A,N140D, N140E, N140F, N140G, N140H, N140I, N140K, N140L, N140M, N140P,N140Q, N140R, N140S, N140T, N140V, N140W, N140Y, G141A, G141D, G141E,G141F, G141H, G141I, G141K, G141L, G141M, G141N, G141P, G141Q, G141R,G141S, G141T, G141V, G141W, G141Y, F142A, F142D, F142E, F142G, F142H,F142I, F142K, F142L, F142M, F142N, F142P, F142Q, F142R, F142S, F142T,F142V, F142W, F142Y, Y143A, Y143D, Y143E, Y143F, Y143G, Y143H, Y143I,Y143K, Y143L, Y143M, Y143N, Y143P, Y143Q, Y143R, Y143S, Y143T, Y143V,Y143W, S144A, S144D, S144E, S144F, S144G, S144H, S144I, S144K, S144L,S144M, S144N, S144P, S144Q, S144R, S144T, S144V, S144W, S144Y, A145D,A145E, A145F, A145G, A145H, A145I, A145K, A145L, A145M, A145N, A145P,A145Q, A145R, A145S, A145T, A145V, A145W, A145Y, N146A, N146D, N146E,N146F, N146G, N146H, N146I, N146K, N146L, N146M, N146P, N146Q, N146R,N146S, N146T, N146V, N146W, N146Y, K147A, K147D, K147E, K147F, K147G,K147H, K147I, K147L, K147M, K147N, K147P, K147Q, K147R, K147S, K147T,K147V, K147W, K147Y, V148A, V148D, V148E, V148F, V148G, V148H, V148I,V148K, V148L, V148M, V148N, V148P, V148Q, V148R, V148S, V148T, V148W,V148Y, A149D, A149E, A149F, A149G, A149H, A149I, A149K, A149L, A149M,A149N, A149P, A149Q, A149R, A149S, A149T, A149V, A149W, A149Y, Q150A,Q150D, Q150E, Q150F, Q150G, Q150H, Q150I, Q150K, Q150L, Q150M, Q150N,Q150P, Q150R, Q150S, Q150T, Q150V, Q150W, Q150Y, F151A, F151D, F151E,F151G, F151H, F151I, F151K, F151L, F151M, F151N, F151P, F151Q, F151R,F151S, F151T, F151V, F151W, F151Y, D152A, D152E, D152F, D152G, D152H,D152I, D152K, D152L, D152M, D152N, D152P, D152Q, D152R, D152S, D152T,D152V, D152W, D152Y, P153A, P153D, P153E, P153F, P153G, P153H, P153I,P153K, P153L, P153M, P153N, P153Q, P153R, P153S, P153T, P153V, P153W,P153Y, S154A, S154D, S154E, S154F, S154G, S154H, S154I, S154K, S154L,S154M, S154N, S154P, S154Q, S154R, S154T, S154V, S154W, S154Y, K155A,K155D, K155E, K155F, K155G, K155H, K155I, K155L, K155M, K155N, K155P,K155Q, K155R, K155S, K155T, K155V, K155W, K155Y, P156A, P156D, P156E,P156F, P156G, P156H, P156I, P156K, P156L, P156M, P156N, P156Q, P156R,P156S, P156T, P156V, P156W, P156Y, Q157A, Q157D, Q157E, Q157F, Q157G,Q157H, Q157I, Q157K, Q157L, Q157M, Q157N, Q157P, Q157R, Q157S, Q157T,Q157V, Q157W, Q157Y, Q158A, Q158D, Q158E, Q158F, Q158G, Q158H, Q158I,Q158K, Q158L, Q158M, Q158N, Q158P, Q158R, Q158S, Q158T, Q158V, Q158W,Q158Y, T159A, T159D, T159E, T159F, T159G, T159H, T159I, T159K, T159L,T159M, T159N, T159P, T159Q, T159R, T159S, T159V, T159W, T159Y, K160A,K160D, K160E, K160F, K160G, K160H, K160I, K160L, K160M, K160N, K160P,K160Q, K160R, K160S, K160T, K160V, K160W, K160Y, G161A, G161D, G161E,G161F, G161H, G161I, G161K, G161L, G161M, G161N, G161P, G161Q, G161R,G161S, G161T, G161V, G161W, G161Y, T162A, T162D, T162E, T162F, T162G,T162H, T162I, T162K, T162L, T162M, T162N, T162P, T162Q, T162R, T162S,T162T, T162V, T162W, T162Y, W163A, W163D, W163E, W163F, W163G, W163H,W163I, W163K, W163L, W163M, W163N, W163P, W163Q, W163R, W163S, W163T,W163V, W163Y, F164A, F164D, F164E, F164G, F164H, F164I, F164K, F164L,F164M, F164N, F164P, F164Q, F164R, F164S, F164T, F164V, F164W, F164Y,Q165A, Q165D, Q165E, Q165F, Q165G, Q165H, Q165I, Q165K, Q165L, Q165M,Q165N, Q165P, Q165R, Q165S, Q165T, Q165V, Q165W, Q165Y, I166A, I166D,I166E, I166F, I166G, I166H, I166K, I166L, I166M, I166N, I166P, I166Q,I166R, I166S, I166T, I166V, I166W, I166Y, T167A, T167D, T167E, T167F,T167G, T167H, T167I, T167K, T167L, T167M, T167N, T167P, T167Q, T167R,T167S, T167V, T167W, T167Y, K168A, K168D, K168E, K168F, K168G, K168H,K168I, K168L, K168M, K168N, K168P, K168Q, K168R, K168S, K168T, K168V,K168W, K168Y, F169A, F169D, F169E, F169G, F169H, F169I, F169K, F169L,F169M, F169N, F169P, F169Q, F169R, F169S, F169T, F169V, F169W, F169Y,T170A, T170D, T170E, T170F, T170G, T170H, T170I, T170K, T170L, T170M,T170N, T170P, T170Q, T170R, T170S, T170V, T170W, T170Y, G171A, G171D,G171E, G171F, G171H, G171I, G171K, G171L, G171M, G171N, G171P, G171Q,G171R, G171S, G171T, G171V, G171W, G171Y, A172D, A172E, A172F, A172G,A172H, A172I, A172K, A172L, A172M, A172N, A172P, A172Q, A172R, A172S,A172T, A172V, A172W, A172Y, A173D, A173E, A173F, A173G, A173H, A173I,A173K, A173L, A173M, A173N, A173P, A173Q, A173R, A173S, A173T, A173V,A173W, A173Y, G174A, G174D, G174E, G174F, G174H, G174I, G174K, G174L,G174M, G174N, G174P, G174Q, G174R, G174S, G174T, G174V, G174W, G174Y,P175A, P175D, P175E, P175F, P175G, P175H, P175I, P175K, P175L, P175M,P175N, P175Q, P175R, P175S, P175T, P175V, P175W, P175Y, Y176A, Y176D,Y176E, Y176F, Y176G, Y176H, Y176I, Y176K, Y176L, Y176M, Y176N, Y176P,Y176Q, Y176R, Y176S, Y176T, Y176V, Y176W, K178A, K178D, K178E, K178F,K178G, K178H, K178I, K178L, K178M, K178N, K178P, K178Q, K178R, K178S,K178T, K178V, K178W, K178Y, A179D, A179E, A179F, A179G, A179H, A179I,A179K, A179L, A179M, A179N, A179P, A179Q, A179R, A179S, A179T, A179V,A179W, A179Y, L180A, L180D, L180E, L180F, L180G, L180H, L180I, L180K,L180M, L180N, L180P, L180Q, L180R, L180S, L180T, L180V, L180W, L180Y,G181A, G181D, G181E, G181F, G181H, G181I, G181K, G181L, G181M, G181N,G181P, G181Q, G181R, G181S, G181T, G181V, G181W, G181Y, S182A, S182D,S182E, S182F, S182G, S182H, S182I, S182K, S182L, S182M, S182N, S182P,S182Q, S182R, S182T, S182V, S182W, S182Y, N183A, N183D, N183E, N183F,N183G, N183H, N183I, N183K, N183L, N183M, N183P, N183Q, N183R, N183S,N183T, N183V, N183W, N183Y, D184A, D184E, D184F, D184G, D184H, D184I,D184K, D184L, D184M, D184N, D184P, D184Q, D184R, D184S, D184T, D184V,D184W, D184Y, K185A, K185D, K185E, K185F, K185G·K185H, K185I, K185L,K185M, K185N, K185P, K185Q, K185R, K185S, K185T, K185V, K185W, K185Y,S186A, S186D, S186E, S186F, S186G, S186H, S186I, S186K, S186L, S186M,S186N, S186P, S186Q, S186R, S186T, S186V, S186W, S186Y, V187A, V187D,V187E, V187F, V187G, V187H, V187I, V187K, V187L, V187M, V187N, V187P,V187Q, V187R, V187S, V187T, V187W, V187Y, D189A, D189E, D189F, D189G,D189H, D189I, D189K, D189L, D189M, D189N, D189P, D189Q, D189R, D189S,D189T, D189V, D189W, D189Y, K190A, K190D, K190E, K190F, K190G, K190H,K190I, K190L, K190M, K190N, K190P, K190Q, K190R, K190S, K190T, K190V,K190W, K190Y, N191A, N191D, N191E, N191F, N191G, N191H, N191I, N191K,N191L, N191M, N191P, N191Q, N191R, N191S, N191T, N191V, N191W, N191Y,K192A, K192D, K192E, K192F, K192G, K192H, K192I, K192L, K192M, K192N,K192P, K192Q, K192R, K192S, K192T, K192V, K192W, K192Y, N193A, N193D,N193E, N193F, N193G, N193H, N193I, N193K, N193L, N193M, N193P, N193Q,N193R, N193S, N193T, N193V, N193W, N193Y, I194A, I194D, I194E, I194F,I194G, I194H, I194K, I194L, I194M, I194N, I194P, I194Q, I194R, I194S,I194T, I194V, I194W, I194Y, A195D, A195E, A195F, A195G, A195H, A195I,A195K, A195L, A195M, A195N, A195P, A195Q, A195R, A195S, A195T, A195V,A195W, A195Y, G196A, G196D, G196E, G196F, G196H, G196I, G196K, G196L,G196M, G196N, G196P, G196Q, G196R, G196S, G196T, G196V, G196W, G196Y,D197A, D197E, D197F, D197G, D197H, D197I, D197K, D197L, D197M, D197N,D197P, D197Q, D197R, D197S, D197T, D197V, D197W, D197Y, W198A, W198D,W198E, W198F, W198G, W198H, W198I, W198K, W198L, W198M, W198N, W198P,W198Q, W198R, W198S, W198T, W198V, W198Y, G199A, G199D, G199E, G199F,G199H, G199I, G199K, G199L, G199M, G199N, G199P, G199Q, G199R, G199S,G199T, G199V, G199W, G199Y, F200A, F200D, F200E, F200G, F200H, F200I,F200K, F200L, F200M, F200N, F200P, F200Q, F200R, F200S, F200T, F200V,F200W, F200Y, D201A, D201E, D201F, D201G, D201H, D201I, D201K, D201L,D201M, D201N, D201P, D201Q, D201R, D201S, D201T, D201V, D201W, D201Y,P202A, P202D, P202E, P202F, P202G, P202H, P202I, P202K, P202L, P202M,P202N, P202Q, P202R, P202S, P202T, P202V, P202W, P202Y, A203D, A203E,A203F, A203G, A203H, A203I, A203K, A203L, A203M, A203N, A203P, A203Q,A203R, A203S, A203T, A203V, A203W, A203Y, K204A, K204D, K204E, K204F,K204G, K204H, K204I, K204L, K204M, K204N, K204P, K204Q, K204R, K204S,K204T, K204V, K204W, K204Y, W205A, W205D, W205E, W205F, W205G, W205H,W205I, W205K, W205L, W205M, W205N, W205P, W205Q, W205R, W205S, W205T,W205V, W205Y, A206D, A206E, A206F, A206G, A206H, A206I, A206K, A206L,A206M, A206N, A206P, A206Q, A206R, A206S, A206T, A206V, A206W, A206Y,Y207A, Y207D, Y207E, Y207F, Y207G, Y207H, Y207I, Y207K, Y207L, Y207M,Y207N, Y207P, Y207Q, Y207R, Y207S, Y207T, Y207V, Y207W, Q208A, Q208D,Q208E, Q208F, Q208G, Q208H, Q208I, Q208K, Q208L, Q208M, Q208N, Q208P,Q208R, Q208S, Q208T, Q208V, Q208W, Q208Y, Y209A, Y209D, Y209E, Y209F,Y209G, Y209H, Y209I, Y209K, Y209L, Y209M, Y209N, Y209P, Y209Q, Y209R,Y209S, Y209T, Y209V, Y209W, D210A, D210E, D210F, D210G, D210H, D210I,D210K, D210L, D210M, D210N, D210P, D210Q, D210R, D210S, D210T, D210V,D210W, D210Y, E211A, E211D, E211F, E211G, E211H, E211I, E211K, E211L,E211M, E211N, E211P, E211Q, E211R, E211S, E211T, E211V, E211W, E211Y,K212A, K212D, K212E, K212F, K212G, K212H, K212I, K212L, K212M, K212N,K212P, K212Q, K212R, K212S, K212T, K212V, K212W, K212Y, N213A, N213D,N213E, N213F, N213G, N213H, N213I, N213K, N213L, N213M, N213P, N213Q,N213R, N213S, N213T, N213V, N213W, N213Y, N214A, N214D, N214E, N214F,N214G, N214H, N214I, N214K, N214L, N214M, N214P, N214Q, N214R, N214S,N214T, N214V, N214W, N214Y, K215A, K215D, K215E, K215F, K215G, K215H,K215I, K215L, K215M, K215N, K215P, K215Q, K215R, K215S, K215T, K215V,K215W, K215Y, F216A, F216D, F216E, F216G, F216H, F216I, F216K, F216L,F216M, F216N, F216P, F216Q, F216R, F216S, F216T, F216V, F216W, F216Y,N217A, N217D, N217E, N217F, N217G, N217H, N217I, N217K, N217L, N217M,N217P, N217Q, N217R, N217S, N217T, N217V, N217W, N217Y, Y218A, Y218D,Y218E, Y218F, Y218G, Y218H, Y218I, Y218K, Y218L, Y218M, Y218N, Y218P,Y218Q, Y218R, Y218S, Y218T, Y218V, Y218W, V219A, V219D, V219E, V219F,V219G, V219H, V219I, V219K, V219L, V219M, V219N, V219P, V219Q, V219R,V219S, V219T, V219W, V219Y, G220A, G220D, G220E, G220F, G220H, G220I,G220K, G220L, G220M, G220N, G220P, G220Q, G220R, G220S, G220T, G220V,G220W, G220Y, K221A, K221D, K221E, K221F, K221G, K221H, K221I, K221L,K221M, K221N, K221P, K221Q, K221R, K221S, K221T, K221V, K221W and K221Y,wherein each position corresponds to the position of the polypeptide ofSEQ ID NO 1, and wherein the variant having improved stability ascompared to the reference DNase e.g. SEQ ID NO 1 i.e. IF>1.0 whenmeasured in the stability assay as described in Example 2.

One embodiment of the invention relates to DNase variants having atleast 80% sequence identity to the polypeptide shown in SEQ ID NO: 1,having DNase activity and comprising one or more substitutions selectedfrom the group consisting of N4E, L17E, T19A, T19G, T19I, K36P, Q38P,S39V, S39R, A40P, A40H, L41T, L41H, V45H, L51G, K53T, K53P, G54P, A55P,N57H, E64A, E64Q, E64R, E64T, E64I, E64S, T66H, K67A, K67T, N68V, N68P,N68I, N68H, S69A, S69D, S69E, S69K, S69L, S69W, S69Y, S69Q, N70T, N70H,N70G, R71T, D72E, S74H, S74G, G75I, N77T, K82P, K82I, D83T, D83P, D83I,D83H, D83G, P84H, Q85T, Q85P, Q85H, K86T, K86P, K86H, G88P, G88H, A91P,W99T, A101W, K105E, K105N, K105T, K105D, S106T, S115T, L116I, Q135L,G136L, V138I, V138L, V138P, V138Q, L139A, N140R, N140L, N140A, G141L,F151R, D152Y, D152L, D152I, D152A, P153E, S154R, T162R, W163E, F164R,I166Y, I166R, K168N, F169R, F169E, A173I, A173R, A173T, S182R, N183E,D184I, K185Y, S186I, D189G, D189H, K212G, K212P and K215I, wherein eachposition corresponds to the position of the polypeptide of SEQ ID NO 1,and wherein the variant having improved stability as compared to thereference DNase e.g. SEQ ID NO 1 i.e. IF>1.0 when measured in thestability assay as described in Example 2.

The variants according to the invention may have improved stabilityand/or also have improved deep cleaning performance. Thus, in apreferred embodiment the variants according to the invention haveimproved detergent stability and/or improved deep cleaning performancecompared to a DNase with SEQ ID NO: 1. In a preferred embodiment, theDNase variant comprises a substitution at one or more positions selectedfrom: N4E, L17E, T19A, T19G, T19I, K36P, Q38P, S39V, S39R, A40P, A40H,L41T, L41H, V45H, L51G, K53T, K53P, G54P, A55P, N57H, E64A, E64Q, E64R,E64T, E64I, E64S, T66H, K67A, K67T, N68V, N68P, N68I, N68H, S69A, S69D,S69E, S69K, S69L, S69W, S69Y, S69Q, N70T, N70H, N70G, R71T, D72E, S74H,S74G, G75I, N77T, K82P, K82I, D83T, D83P, D83I, D83H, D83G, P84H, Q85T,Q85P, Q85H, K86T, K86P, K86H, G88P, G88H, A91P, W99T, A101W, K105E,K105N, K105T, K105D, S106T, S115T, L116I, Q135L, G136L, V138I, V138L,V138P, V138Q, L139A, N140R, N140L, N140A, G141L, F151R, D152Y, D152L,D152I, D152A, P153E, S154R, T162R, W163E, F164R, I166Y, I166R, K168N,F169R, F169E, A173I, A173R, A173T, S182R, N183E, D184I, K185Y, S186I,D189G, D189H, K212G, K212P and K215I, wherein each position correspondsto the position of the polypeptide of SEQ ID NO 1 and wherein thevariant has at least 60%, such as at least 61%, at least 62%, at least63%, at least 64%, at least 65%, at least 66%, at least 67%, at least68%, at least 69%, at least 70%, at least 71%, at least 72%, at least73%, at least 74%, at least 75%, at least 76%, at least 77%, at least78%, at least 79%, at least 80%, at least 81%, at least 82%, at least83%, at least 84%, at least 85%, at least 86%, at least 87%, at least88%, at least 89%, at least 90%, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98% or at least 99%, sequence identity to SEQ ID NO: 1.

The variants according to the invention may further comprise one or moreadditional alterations at one or more (e.g., several) other positions.The amino acid changes may be of a minor nature, that is conservativeamino acid substitutions or insertions that do not significantly affectthe folding and/or activity of the protein; small deletions, typicallyof 1-5 amino acids; small amino- or carboxyl-terminal extensions, suchas an amino-terminal methionine residue; a small linker peptide of up to20-25 residues, located at the amino- or carboxyl terminal; or a smallextension that facilitates purification by changing net charge oranother function, such as a poly-histidine tract, an antigenic epitopeor a binding domain.

Examples of conservative substitutions are within the groups of basicamino acids (arginine, lysine and histidine), acidic amino acids(glutamic acid and aspartic acid), polar amino acids (glutamine andasparagine), hydrophobic amino acids (leucine, isoleucine and valine),aromatic amino acids (phenylalanine, tryptophan and tyrosine), and smallamino acids (glycine, alanine, serine, threonine and methionine). Aminoacid substitutions that do not generally alter specific activity areknown in the art and are described, for example, by H. Neurath and R. L.Hill, 1979, In, The Proteins, Academic Press, New York. Commonsubstitutions are Ala/Ser, Val/Ile, Asp/Glu, Asn/Gln, Thr/Ser, Ala/Gly,Ala/Thr, Ser/Asn, Ala/Val, Ser/Gly, Tyr/Phe, Ala/Pro, Lys/Arg, Asp/Asn,Glu/Gln, Leu/Ile, Leu/Val, Ala/Glu, and Asp/Gly.

Alternatively, the amino acid changes are of such a nature that thephysico-chemical properties of the polypeptides are altered. Forexample, amino acid changes may improve the thermal stability of thepolypeptide, alter the substrate specificity, change the pH optimum, andthe like.

Essential amino acids in a polypeptide can be identified according toprocedures known in the art, such as site-directed mutagenesis oralanine-scanning mutagenesis (Cunningham and Wells, 1989, Science 244:1081-1085). In the latter technique, single alanine mutations areintroduced at every residue in the molecule, and the resultant mutantmolecules are tested for DNase activity to identify amino acid residuesthat are critical to the activity of the molecule. See also, Hilton etal., 1996, J. Biol. Chem. 271: 4699-4708. The active site of the enzymeor other biological interaction can also be determined by physicalanalysis of structure, as determined by such techniques as nuclearmagnetic resonance, crystallography, electron diffraction, orphotoaffinity labeling, in conjunction with mutation of putative contactsite amino acids. See, for example, de Vos et al., 1992, Science 255:306-312; Smith et al., 1992, J. Mol. Biol. 224: 899-904; Wlodaver etal., 1992, FEBS Lett. 309: 59-64. The identity of essential amino acidscan also be inferred from an alignment with a related polypeptide.

Substantially DNases variants may have one or more (several) amino acidsubstitutions, deletions and/or insertions as described above. In thepresent context the term “one or more” is used interchangeably with theterm “several”. The DNase variants of the invention may further comprisechanges which preferably are of a minor nature, e.g. conservative aminoacid substitutions and other substitutions that do not significantlyaffect the three-dimensional folding or activity of the protein orpolypeptide; small deletions, typically of one to about 30 amino acids;and small amino- or carboxyl-terminal extensions, such as anamino-terminal methionine residue, a small linker peptide of up to about20-25 residues, or a small extension that facilitates purification (anaffinity tag), such as a poly-histidine tract, or protein A (Nilsson etal., 1985, EMBO J. 4: 1075; Nilsson et al., 1991, Methods Enzymol. 198:3. See, also, in general, Ford et al., 1991, Protein Expression andPurification 2: 95-107.

Although the changes described above preferably are of a minor nature,such changes may also be of a substantive nature such as fusion oflarger polypeptides of up to 300 amino acids or more both as amino- orcarboxyl-terminal extensions.

The parent DNase may comprise or consist of the amino acid sequence ofSEQ ID NO: 1 or an allelic variant thereof; or a fragment thereof havingDNase activity. In one aspect, the parent DNase comprises or consists ofthe amino acid sequence of SEQ ID NO: 1.

In a preferred aspect of the invention the parent DNase comprise apolypeptide having at least 60%, such as at least 61%, at least 62%, atleast 63%, at least 64%, at least 65%, at least 66%, at least 67%, atleast 68%, at least 69%, at least 70%, at least 71%, at least 72%, atleast 73%, at least 74%, at least 75%, at least 76%, at least 77%, atleast 78%, at least 79%, at least 80%, at least 81%, at least 82%, atleast 83%, at least 84%, at least 85%, at least 86%, at least 87%, atleast 88%, at least 89%, at least 90%, at least 91%, at least 92%, atleast 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, at least 99%, or 100% sequence identity to the polypeptideshown in SEQ ID NO: 1.

The parent DNase may be (a) a polypeptide having at least 60% sequenceidentity to the polypeptide of SEQ ID NO: 1; (b) a polypeptide encodedby a polynucleotide that hybridizes under medium or high stringencyconditions with (i) the mature polypeptide coding sequence of SEQ ID NO:1, (ii) the cDNA sequence thereof, or (iii) the full-length complementof (i) or (ii); or (c).

In another aspect, the parent DNase is encoded by a polynucleotide thathybridizes under very low stringency conditions, low stringencyconditions, medium stringency conditions, or high stringency conditions,or very high stringency conditions with (i) the mature polypeptidecoding sequence, (ii) the cDNA sequence thereof, or (iii) thefull-length complement of (i) or (ii) (Sambrook et al., 1989, MolecularCloning, A Laboratory Manual, 2d edition, Cold Spring Harbor, N.Y.).

The polynucleotide or a subsequence thereof, as well as the polypeptideof SEQ ID NO: 1 or a fragment thereof may be used to design nucleic acidprobes to identify and clone DNA encoding a parent from strains ofdifferent genera or species according to methods well known in the art.In particular, such probes can be used for hybridization with thegenomic DNA or cDNA of a cell of interest, following standard Southernblotting procedures, in order to identify and isolate the correspondinggene therein. Such probes can be considerably shorter than the entiresequence, but should be at least 15, e.g., at least 25, at least 35, orat least 70 nucleotides in length. Preferably, the nucleic acid probe isat least 100 nucleotides in length, e.g., at least 200 nucleotides, atleast 300 nucleotides, at least 400 nucleotides, at least 500nucleotides or at least 600 nucleotides in length. Both DNA and RNAprobes can be used. The probes are typically labeled for detecting thecorresponding gene (for example, with ³²P, ³H, ³⁵S, biotin, or avidin).Such probes are encompassed by the present invention.

A genomic DNA or cDNA library prepared from such other strains may bescreened for DNA that hybridizes with the probes described above andencodes a polypeptide having DNase activity. Genomic or other DNA fromsuch other strains may be separated by agarose or polyacrylamide gelelectrophoresis, or other separation techniques. DNA from the librariesor the separated DNA may be transferred to and immobilized onnitrocellulose or other suitable carrier material. In order to identifya clone or DNA that hybridizes with a polynucleotide encoding SEQ ID NO:1 or a subsequence thereof, the carrier material is used in a Southernblot.

For purposes of the present invention, hybridization indicates that thepolynucleotide hybridizes to a labeled nucleic acid probe correspondingto (i) the mature polypeptide coding sequence; (ii) the cDNA sequencethereof; (iii) the full-length complement thereof; or (iv) a subsequencethereof; under very low, low stringency conditions, low-mediumstringency conditions, medium stringency conditions, medium-highstringency conditions, high stringency conditions to very highstringency conditions. Molecules to which the nucleic acid probehybridizes under these conditions can be detected using, for example, Xray film or any other detection means known in the art.

The parent DNase may be obtained from organisms of any genus. Forpurposes of the present invention, the term “obtained from” as usedherein in connection with a given source shall mean that the parentencoded by a polynucleotide is produced by the source or by a strain inwhich the polynucleotide from the source has been inserted. In oneaspect, the parent is secreted extracellularly.

The parent may be a fungal DNase. For example, the parent may be anAspergillus DNase. In one aspect, the parent is an Aspergillus oryzaeDNase, e.g., a DNase with SEQ ID NO: 1.

Strains of these species are readily accessible to the public in anumber of culture collections, such as the American Type CultureCollection (ATCC), Deutsche Sammlung von Mikroorganismen undZellkulturen GmbH (DSMZ), Centraalbureau Voor Schimmelcultures (CBS),and Agricultural Research Service Patent Culture Collection, NorthernRegional Research Center (NRRL).

The parent may be identified and obtained from other sources includingmicroorganisms isolated from nature (e.g., soil, composts, water, etc.)or DNA samples obtained directly from natural materials (e.g., soil,composts, water, etc.) using the above-mentioned probes. Techniques forisolating microorganisms and DNA directly from natural habitats are wellknown in the art. A polynucleotide encoding a parent may then beobtained by similarly screening a genomic DNA or cDNA library of anothermicroorganism or mixed DNA sample. Once a polynucleotide encoding aparent has been detected with the probe(s), the polynucleotide can beisolated or cloned by utilizing techniques that are known to those ofordinary skill in the art (see, e.g., Sambrook et al., 1989, supra).

Preparation of Variants

The present invention also relates to a method for obtaining a DNasevariant having at least one improved property compared to SEQ ID NO: 1,comprising

a) introducing into a parent DNase with at least 60% identity to of SEQID NO: 1 an alteration at one or more positions selected from: 1, 2, 3,4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22,23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40,41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 53, 54, 55, 56, 57, 58, 59,60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77,78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95,96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110,111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124,125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138,139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152,153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166,167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180,181, 182, 183, 185, 186, 187, 189, 190, 191, 192, 193, 194, 195, 196,197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210,211, 212, 213, 214, 215, 216, 217, 218, 219, 220 and 221, wherein thevariant has an amino acid sequence which is at least 60%, at least 70%,at least 80%, at least 85%, at least 90% or at least 95% identical toSEQ ID NO: 1; and

b) recovering the variant.

The present invention relates to a method for obtaining a DNase varianthaving at least one improved property compared to SEQ ID NO: 1,comprising introducing into a parent DNase with at least 60% identity toSEQ ID NO: 1 an alteration at one or more positions: 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25,26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43,44, 45, 46, 47, 48, 49, 50, 51, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62,63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80,81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98,99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112,113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126,127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140,141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154,155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168,169, 170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182,183, 185, 186, 187, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198,199, 200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212,213, 214, 215, 216, 217, 218, 219, 220 or 221, wherein the variant hasan amino acid sequence which is at least 60%, at least 70%, at least80%, at least 90% or at least 95% identical to SEQ ID NO: 1; andrecovering the variant.

The present invention also relates to a method for obtaining a DNasevariant having at least one improved property compared to SEQ ID NO: 1,comprising

a) introducing into a parent DNase with at least 80% identity to of SEQID NO: 1 an alteration at one or more positions selected from: N4E,L17E, T19A, T19G, T19I, K36P, Q38P, S39V, S39R, A40P, A40H, L41T, L41H,V45H, L51G, K53T, K53P, G54P, A55P, N57H, E64A, E64Q, E64R, E64T, E64I,E64S, T66H, K67A, K67T, N68V, N68P, N68I, N68H, S69A, S69D, S69E, S69K,S69L, S69W, S69Y, S69Q, N70T, N70H, N70G, R71T, D72E, S74H, S74G, G75I,N77T, K82P, K82I, D83T, D83P, D83I, D83H, D83G, P84H, Q85T, Q85P, Q85H,K86T, K86P, K86H, G88P, G88H, A91P, W99T, A101W, K105E, K105N, K105T,K105D, S106T, S115T, L116I, Q135L, G136L, V138I, V138L, V138P, V138Q,L139A, N140R, N140L, N140A, G141L, F151R, D152Y, D152L, D152I, D152A,P153E, S154R, T162R, W163E, F164R, I166Y, I166R, K168N, F169R, F169E,A173I, A173R, A173T, S182R, N183E, D184I, K185Y, S186I, D189G, D189H,K212G, K212P and K215I, wherein the variant has an amino acid sequencewhich is at least 60%, at least 70%, at least 80%, at least 85%, atleast 90% or at least 95% identical to SEQ ID NO: 1; and

b) recovering the variant.

The variants of the invention may also be prepared by procedures such asthose mentioned below.

Site-directed mutagenesis is a technique in which one or more (e.g.,several) mutations are introduced at one or more defined sites in apolynucleotide encoding the parent.

Site-directed mutagenesis can be accomplished in vitro by PCR involvingthe use of oligonucleotide primers containing the desired mutation.Site-directed mutagenesis can also be performed in vitro by cassettemutagenesis involving the cleavage by a restriction enzyme at a site inthe plasmid comprising a polynucleotide encoding the parent andsubsequent ligation of an oligonucleotide containing the mutation in thepolynucleotide. Usually the restriction enzyme that digests the plasmidand the oligonucleotide is the same, permitting sticky ends of theplasmid and the insert to ligate to one another. See, e.g., Scherer andDavis, 1979, Proc. Natl. Acad. Sci. USA 76: 4949-4955; and Barton etal., 1990, Nucleic Acids Res. 18: 7349-4966.

Site-directed mutagenesis can also be accomplished in vivo by methodsknown in the art. See, e.g., U.S. Patent Application Publication No.2004/0171154; Storici et al., 2001, Nature Biotechnol. 19: 773-776; Krenet al., 1998, Nat. Med. 4: 285-290; and Calissano and Macino, 1996,Fungal Genet. Newslett. 43: 15-16.

Synthetic gene construction entails in vitro synthesis of a designedpolynucleotide molecule to encode a polypeptide of interest. Genesynthesis can be performed utilizing a number of techniques, such as themultiplex microchip-based technology described by Tian et al. (2004,Nature 432: 1050-1054) and similar technologies wherein oligonucleotidesare synthesized and assembled upon photo-programmable microfluidicchips.

Single or multiple amino acid substitutions, deletions, and/orinsertions can be made and tested using known methods of mutagenesis,recombination, and/or shuffling, followed by a relevant screeningprocedure, such as those disclosed by Reidhaar-Olson and Sauer, 1988,Science 241: 53-57; Bowie and Sauer, 1989, Proc. Natl. Acad. Sci. USA86: 2152-2156; WO 95/17413; or WO 95/22625. Other methods that can beused include error-prone PCR, phage display (e.g., Lowman et al., 1991,Biochemistry 30: 10832-10837; U.S. Pat. No. 5,223,409; WO 92/06204) andregion-directed mutagenesis (Derbyshire et al., 1986, Gene 46: 145; Neret al., 1988, DNA 7: 127).

Mutagenesis/shuffling methods can be combined with high-throughput,automated screening methods to detect activity of cloned, mutagenizedpolypeptides expressed by host cells (Ness et al., 1999, NatureBiotechnology 17: 893-896). Mutagenized DNA molecules that encode activepolypeptides can be recovered from the host cells and rapidly sequencedusing standard methods in the art. These methods allow the rapiddetermination of the importance of individual amino acid residues in apolypeptide.

Semi-synthetic gene construction is accomplished by combining aspects ofsynthetic gene construction, and/or site-directed mutagenesis, and/orrandom mutagenesis, and/or shuffling. Semi-synthetic construction istypified by a process utilizing polynucleotide fragments that aresynthesized, in combination with PCR techniques. Defined regions ofgenes may thus be synthesized de novo, while other regions may beamplified using site-specific mutagenic primers, while yet other regionsmay be subjected to error-prone PCR or non-error prone PCRamplification. Polynucleotide subsequences may then be shuffled.

Nucleic Acid Constructs

The present invention also relates to nucleic acid constructs comprisinga polynucleotide of the present invention operably linked to one or morecontrol sequences that direct the expression of the coding sequence in asuitable host cell under conditions compatible with the controlsequences.

The polynucleotide may be manipulated in a variety of ways to providefor expression of the polypeptide. Manipulation of the polynucleotideprior to its insertion into a vector may be desirable or necessarydepending on the expression vector. The techniques for modifyingpolynucleotides utilizing recombinant DNA methods are well known in theart.

The control sequence may be a promoter, a polynucleotide that isrecognized by a host cell for expression of a polynucleotide encoding apolypeptide of the present invention. The promoter containstranscriptional control sequences that mediate the expression of thepolypeptide. The promoter may be any polynucleotide that showstranscriptional activity in the host cell including mutant, truncated,and hybrid promoters, and may be obtained from genes encodingextracellular or intracellular polypeptides either homologous orheterologous to the host cell.

Examples of suitable promoters for directing transcription of thenucleic acid constructs of the present invention in a bacterial hostcell are the promoters obtained from the Bacillus amyloliquefaciensalpha-amylase gene (amyQ), Bacillus licheniformis alpha-amylase gene(amyL), Bacillus licheniformis penicillinase gene (penP), Bacillusstearothermophilus maltogenic amylase gene (amyM), Bacillus subtilislevansucrase gene (sacB), Bacillus subtilis xylA and xylB genes,Bacillus thuringiensis cryIIIA gene (Agaisse and Lereclus, 1994,Molecular Microbiology 13: 97-107), E. coli lac operon, E. coli trcpromoter (Egon et al., 1988, Gene 69: 301-315), Streptomyces coelicoloragarase gene (dagA), and prokaryotic beta-lactamase gene (Villa-Kamaroffet al., 1978, Proc. Natl. Acad. Sci. USA 75: 3727-3731), as well as thetac promoter (DeBoer et al., 1983, Proc. Natl. Acad. Sci. USA 80:21-25). Further promoters are described in “Useful proteins fromrecombinant bacteria” in Gilbert et al., 1980, Scientific American 242:74-94; and in Sambrook et al., 1989, supra. Examples of tandem promotersare disclosed in WO 99/43835.

Examples of suitable promoters for directing transcription of thenucleic acid constructs of the present invention in a filamentous fungalhost cell are promoters obtained from the genes for Aspergillus nidulansacetamidase, Aspergillus niger neutral alpha-amylase, Aspergillus nigeracid stable alpha-amylase, Aspergillus niger or Aspergillus awamoriglucoamylase (glaA), Aspergillus oryzae TAKA amylase, Aspergillus oryzaealkaline protease, Aspergillus oryzae triose phosphate isomerase,Fusarium oxysporum trypsin-like protease (WO 96/00787), Fusariumvenenatum amyloglucosidase (WO 00/56900), Fusarium venenatum Daria (WO00/56900), Fusarium venenatum Quinn (WO 00/56900), Rhizomucor mieheilipase, Rhizomucor miehei aspartic proteinase, Trichoderma reeseibeta-glucosidase, Trichoderma reesei cellobiohydrolase I, Trichodermareesei cellobiohydrolase II, Trichoderma reesei endoglucanase I,Trichoderma reesei endoglucanase II, Trichoderma reesei endoglucanaseIll, Trichoderma reesei endoglucanase V, Trichoderma reesei xylanase I,Trichoderma reesei xylanase II, Trichoderma reesei xylanase Ill,Trichoderma reesei beta-xylosidase, and Trichoderma reesei translationelongation factor, as well as the NA2 tpi promoter (a modified promoterfrom an Aspergillus neutral alpha-amylase gene in which the untranslatedleader has been replaced by an untranslated leader from an Aspergillustriose phosphate isomerase gene; non-limiting examples include modifiedpromoters from an Aspergillus niger neutral alpha-amylase gene in whichthe untranslated leader has been replaced by an untranslated leader froman Aspergillus nidulans or Aspergillus oryzae triose phosphate isomerasegene); and mutant, truncated, and hybrid promoters thereof. Otherpromoters are described in U.S. Pat. No. 6,011,147.

In a yeast host, useful promoters are obtained from the genes forSaccharomyces cerevisiae enolase (ENO 1), Saccharomyces cerevisiaegalactokinase (GAL1), Saccharomyces cerevisiae alcoholdehydrogenase/glyceraldehyde-3 phosphate dehydrogenase (ADH1, ADH2/GAP),Saccharomyces cerevisiae triose phosphate isomerase (TPI), Saccharomycescerevisiae metallothionein (CUP1), and Saccharomyces cerevisiae 3phosphoglycerate kinase. Other useful promoters for yeast host cells aredescribed by Romanos et al., 1992, Yeast 8: 423-488.

The control sequence may also be a transcription terminator, which isrecognized by a host cell to terminate transcription. The terminator isoperably linked to the 3′ terminus of the polynucleotide encoding thepolypeptide. Any terminator that is functional in the host cell may beused in the present invention.

Preferred terminators for bacterial host cells are obtained from thegenes for Bacillus clausii alkaline protease (aprH), Bacilluslicheniformis alpha-amylase (amyL), and Escherichia coli ribosomal RNA(rrnB).

Preferred terminators for filamentous fungal host cells are obtainedfrom the genes for Aspergillus nidulans acetamidase, Aspergillusnidulans anthranilate synthase, Aspergillus niger glucoamylase,Aspergillus niger alpha-glucosidase, Aspergillus oryzae TAKA amylase,Fusarium oxysporum trypsin-like protease, Trichoderma reeseibeta-glucosidase, Trichoderma reesei cellobiohydrolase I, Trichodermareesei cellobiohydrolase II, Trichoderma reesei endoglucanase I,Trichoderma reesei endoglucanase II, Trichoderma reesei endoglucanaseIll, Trichoderma reesei endoglucanase V, Trichoderma reesei xylanase I,Trichoderma reesei xylanase II, Trichoderma reesei xylanase Ill,Trichoderma reesei beta-xylosidase, and Trichoderma reesei translationelongation factor.

Preferred terminators for yeast host cells are obtained from the genesfor Saccharomyces cerevisiae enolase, Saccharomyces cerevisiaecytochrome C (CYC1), and Saccharomyces cerevisiae glyceraldehyde-3phosphate dehydrogenase. Other useful terminators for yeast host cellsare described by Romanos et al., 1992, supra.

The control sequence may also be an mRNA stabilizer region downstream ofa promoter and upstream of the coding sequence of a gene which increasesexpression of the gene.

Examples of suitable mRNA stabilizer regions are obtained from aBacillus thuringiensis cryIIIA gene (WO 94/25612) and a Bacillussubtilis SP82 gene (Hue et al., 1995, Journal of Bacteriology 177:3465-3471).

The control sequence may also be a leader, a nontranslated region of anmRNA that is important for translation by the host cell. The leader isoperably linked to the 5′ terminus of the polynucleotide encoding thepolypeptide. Any leader that is functional in the host cell may be used.

Preferred leaders for filamentous fungal host cells are obtained fromthe genes for Aspergillus oryzae TAKA amylase and Aspergillus nidulanstriose phosphate isomerase.

Suitable leaders for yeast host cells are obtained from the genes forSaccharomyces cerevisiae enolase (ENO 1), Saccharomyces cerevisiae 3phosphoglycerate kinase, Saccharomyces cerevisiae alpha-factor, andSaccharomyces cerevisiae alcohol dehydrogenase/glyceraldehyde-3phosphate dehydrogenase (ADH2/GAP).

The control sequence may also be a polyadenylation sequence; a sequenceoperably linked to the 3′ terminus of the polynucleotide and, whentranscribed, is recognized by the host cell as a signal to addpolyadenosine residues to transcribed mRNA. Any polyadenylation sequencethat is functional in the host cell may be used.

Preferred polyadenylation sequences for filamentous fungal host cellsare obtained from the genes for Aspergillus nidulans anthranilatesynthase, Aspergillus niger glucoamylase, Aspergillus nigeralpha-glucosidase Aspergillus oryzae TAKA amylase, and Fusariumoxysporum trypsin-like protease.

Useful polyadenylation sequences for yeast host cells are described byGuo and Sherman, 1995, Mol. Cellular Biol. 15: 5983-5990.

The control sequence may also be a signal peptide coding region thatencodes a signal peptide linked to the N terminus of a polypeptide anddirects the polypeptide into the cell's secretory pathway. The 5′ end ofthe coding sequence of the polynucleotide may inherently contain asignal peptide coding sequence naturally linked in translation readingframe with the segment of the coding sequence that encodes thepolypeptide. Alternatively, the 5′ end of the coding sequence maycontain a signal peptide coding sequence that is foreign to the codingsequence. A foreign signal peptide coding sequence may be required wherethe coding sequence does not naturally contain a signal peptide codingsequence. Alternatively, a foreign signal peptide coding sequence maysimply replace the natural signal peptide coding sequence in order toenhance secretion of the polypeptide. However, any signal peptide codingsequence that directs the expressed polypeptide into the secretorypathway of a host cell may be used.

Effective signal peptide coding sequences for bacterial host cells arethe signal peptide coding sequences obtained from the genes for BacillusNCIB 11837 maltogenic amylase, Bacillus licheniformis subtilisin,Bacillus licheniformis beta-lactamase, Bacillus stearothermophilusalpha-amylase, Bacillus stearothermophilus neutral proteases (nprT,nprS, nprM), and Bacillus subtilis prsA. Further signal peptides aredescribed by Simonen and Palva, 1993, Microbiological Reviews 57:109-137.

Effective signal peptide coding sequences for filamentous fungal hostcells are the signal peptide coding sequences obtained from the genesfor Aspergillus niger neutral amylase, Aspergillus niger glucoamylase,Aspergillus oryzae TAKA amylase, Humicola insolens cellulase, Humicolainsolens endoglucanase V, Humicola lanuginosa lipase, and Rhizomucormiehei aspartic proteinase.

Useful signal peptides for yeast host cells are obtained from the genesfor Saccharomyces cerevisiae alpha-factor and Saccharomyces cerevisiaeinvertase. Other useful signal peptide coding sequences are described byRomanos et al., 1992, supra.

The control sequence may also be a propeptide coding sequence thatencodes a propeptide positioned at the N terminus of a polypeptide. Theresultant polypeptide is known as a proenzyme or propolypeptide (or azymogen in some cases). A propolypeptide is generally inactive and canbe converted to an active polypeptide by catalytic or autocatalyticcleavage of the propeptide from the propolypeptide. The propeptidecoding sequence may be obtained from the genes for Bacillus subtilisalkaline protease (aprE), Bacillus subtilis neutral protease (nprT),Myceliophthora thermophila laccase (WO 95/33836), Rhizomucor mieheiaspartic proteinase, and Saccharomyces cerevisiae alpha-factor.

Where both signal peptide and propeptide sequences are present, thepropeptide sequence is positioned next to the N terminus of apolypeptide and the signal peptide sequence is positioned next to the Nterminus of the propeptide sequence.

It may also be desirable to add regulatory sequences that regulateexpression of the polypeptide relative to the growth of the host cell.Examples of regulatory sequences are those that cause expression of thegene to be turned on or off in response to a chemical or physicalstimulus, including the presence of a regulatory compound. Regulatorysequences in prokaryotic systems include the lac, tac, and trp operatorsystems. In yeast, the ADH2 system or GAL1 system may be used. Infilamentous fungi, the Aspergillus niger glucoamylase promoter,Aspergillus oryzae TAKA alpha-amylase promoter, and Aspergillus oryzaeglucoamylase promoter, Trichoderma reesei cellobiohydrolase I promoter,and Trichoderma reesei cellobiohydrolase II promoter may be used. Otherexamples of regulatory sequences are those that allow for geneamplification. In eukaryotic systems, these regulatory sequences includethe dihydrofolate reductase gene that is amplified in the presence ofmethotrexate, and the metallothionein genes that are amplified withheavy metals. In these cases, the polynucleotide encoding thepolypeptide would be operably linked to the regulatory sequence.

Expression Vectors

The present invention also relates to recombinant expression vectorscomprising a polynucleotide of the present invention, a promoter, andtranscriptional and translational stop signals. The various nucleotideand control sequences may be joined together to produce a recombinantexpression vector that may include one or more convenient restrictionsites to allow for insertion or substitution of the polynucleotideencoding the polypeptide at such sites. Alternatively, thepolynucleotide may be expressed by inserting the polynucleotide or anucleic acid construct comprising the polynucleotide into an appropriatevector for expression. In creating the expression vector, the codingsequence is located in the vector so that the coding sequence isoperably linked with the appropriate control sequences for expression.In a further embodiment, polynucleotide sequence codons have beenmodified by nucleotide substitutions to correspond to the codon usage ofthe host organism intended for production of the polypeptide of thepresent invention. The recombinant expression vector may be any vector(e.g., a plasmid or virus) that can be conveniently subjected torecombinant DNA procedures and can bring about expression of thepolynucleotide. The choice of the vector will typically depend on thecompatibility of the vector with the host cell into which the vector isto be introduced. The vector may be a linear or closed circular plasmid.

The vector may be an autonomously replicating vector, i.e., a vectorthat exists as an extrachromosomal entity, the replication of which isindependent of chromosomal replication, e.g., a plasmid, anextrachromosomal element, a minichromosome, or an artificial chromosome.The vector may contain any means for assuring self-replication.Alternatively, the vector may be one that, when introduced into the hostcell, is integrated into the genome and replicated together with thechromosome(s) into which it has been integrated. Furthermore, a singlevector or plasmid or two or more vectors or plasmids that togethercontain the total DNA to be introduced into the genome of the host cell,or a transposon, may be used.

The vector preferably contains one or more selectable markers thatpermit easy selection of transformed, transfected, transduced, or thelike cells. A selectable marker is a gene the product of which providesfor biocide or viral resistance, resistance to heavy metals, prototrophyto auxotrophs, and the like.

Examples of bacterial selectable markers are Bacillus licheniformis orBacillus subtilis dal genes, or markers that confer antibioticresistance such as ampicillin, chloramphenicol, kanamycin, neomycin,spectinomycin, or tetracycline resistance. Suitable markers for yeasthost cells include, but are not limited to, ADE2, HIS3, LEU2, LYS2,MET3, TRP1, and URA3. Selectable markers for use in a filamentous fungalhost cell include, but are not limited to, adeA(phosphoribosylaminoimidazole-succinocarboxamide synthase), adeB(phosphoribosylaminoimidazole synthase), amdS (acetamidase), argB(ornithine carbamoyltransferase), bar (phosphinothricinacetyltransferase), hph (hygromycin phosphotransferase), niaD (nitratereductase), pyrG (orotidine-5′ phosphate decarboxylase), sC (sulfateadenyltransferase), and trpC (anthranilate synthase), as well asequivalents thereof. Preferred for use in an Aspergillus cell areAspergillus nidulans orAspergillus oryzae amdS and pyrG genes and aStreptomyces hygroscopicus bar gene. Preferred for use in a Trichodermacell are adeA, adeB, amdS, hph, and pyrG genes.

The selectable marker may be a dual selectable marker system asdescribed in WO 2010/039889. In one aspect, the dual selectable markeris an hph-tk dual selectable marker system.

The vector preferably contains an element(s) that permits integration ofthe vector into the host cell's genome or autonomous replication of thevector in the cell independent of the genome. For integration into thehost cell genome, the vector may rely on the polynucleotide's sequenceencoding the polypeptide or any other element of the vector forintegration into the genome by homologous or non-homologousrecombination. Alternatively, the vector may contain additionalpolynucleotides for directing integration by homologous recombinationinto the genome of the host cell at a precise location(s) in thechromosome(s). To increase the likelihood of integration at a preciselocation, the integrational elements should contain a sufficient numberof nucleic acids, such as 100 to 10,000 base pairs, 400 to 10,000 basepairs, and 800 to 10,000 base pairs, which have a high degree ofsequence identity to the corresponding target sequence to enhance theprobability of homologous recombination. The integrational elements maybe any sequence that is homologous with the target sequence in thegenome of the host cell. Furthermore, the integrational elements may benon-encoding or encoding polynucleotides. On the other hand, the vectormay be integrated into the genome of the host cell by non-homologousrecombination.

For autonomous replication, the vector may further comprise an origin ofreplication enabling the vector to replicate autonomously in the hostcell in question. The origin of replication may be any plasmidreplicator mediating autonomous replication that functions in a cell.The term “origin of replication” or “plasmid replicator” means apolynucleotide that enables a plasmid or vector to replicate in vivo.

Examples of bacterial origins of replication are the origins ofreplication of plasmids pBR322, pUC19, pACYC177, and pACYC184 permittingreplication in E. coli, and pUB110, pE194, pTA1060, and pAMß1 permittingreplication in Bacillus.

Examples of origins of replication for use in a yeast host cell are the2 micron origin of replication, ARS1, ARS4, the combination of ARS1 andCEN3, and the combination of ARS4 and CEN6.

Examples of origins of replication useful in a filamentous fungal cellare AMA1 and ANS1 (Gems et al., 1991, Gene 98: 61-67; Cullen et al.,1987, Nucleic Acids Res. 15: 9163-9175; WO 00/24883). Isolation of theAMA1 gene and construction of plasmids or vectors comprising the genecan be accomplished according to the methods disclosed in WO 00/24883.

More than one copy of a polynucleotide of the present invention may beinserted into a host cell to increase production of a polypeptide. Anincrease in the copy number of the polynucleotide can be obtained byintegrating at least one additional copy of the sequence into the hostcell genome or by including an amplifiable selectable marker gene withthe polynucleotide where cells containing amplified copies of theselectable marker gene, and thereby additional copies of thepolynucleotide, can be selected for by cultivating the cells in thepresence of the appropriate selectable agent.

The procedures used to ligate the elements described above to constructthe recombinant expression vectors of the present invention are wellknown to one skilled in the art (see, e.g., Sambrook et al., 1989,supra).

Host Cells

The present invention also relates to recombinant host cells, comprisinga polynucleotide of the present invention operably linked to one or morecontrol sequences that direct the production of a polypeptide of thepresent invention. A construct or vector comprising a polynucleotide isintroduced into a host cell so that the construct or vector ismaintained as a chromosomal integrant or as a self-replicatingextra-chromosomal vector as described earlier. The term “host cell”encompasses any progeny of a parent cell that is not identical to theparent cell due to mutations that occur during replication. The choiceof a host cell will to a large extent depend upon the gene encoding thepolypeptide and its source.

The host cell may be any cell useful in the recombinant production of apolypeptide of the present invention, e.g., a prokaryote or a eukaryote.

The prokaryotic host cell may be any Gram-positive or Gram-negativebacterium. Gram-positive bacteria include, but are not limited to,Bacillus, Clostridium, Enterococcus, Geobacillus, Lactobacillus,Lactococcus, Oceanobacillus, Staphylococcus, Streptococcus, andStreptomyces. Gram-negative bacteria include, but are not limited to,Campylobacter, E. coli, Flavobacterium, Fusobacterium, Helicobacter,Ilyobacter, Neisseria, Pseudomonas, Salmonella, and Ureaplasma.

The bacterial host cell may be any Bacillus cell including, but notlimited to, Bacillus alkalophilus, Bacillus amyloliquefaciens, Bacillusbrevis, Bacillus circulans, Bacillus clausii, Bacillus coagulans,Bacillus firmus, Bacillus lautus, Bacillus lentus, Bacilluslicheniformis, Bacillus megaterium, Bacillus pumilus, Bacillusstearothermophilus, Bacillus subtilis, and Bacillus thuringiensis cells.

The bacterial host cell may also be any Streptococcus cell including,but not limited to, Streptococcus equisimilis, Streptococcus pyogenes,Streptococcus uberis, and Streptococcus equi subsp. Zooepidemicus cells.

The bacterial host cell may also be any Streptomyces cell including, butnot limited to, Streptomyces achromogenes, Streptomyces avermitilis,Streptomyces coelicolor, Streptomyces griseus, and Streptomyces lividanscells.

The introduction of DNA into a Bacillus cell may be effected byprotoplast transformation (see, e.g., Chang and Cohen, 1979, Mol. Gen.Genet. 168: 111-115), competent cell transformation (see, e.g., Youngand Spizizen, 1961, J. Bacteriol. 81: 823-829, or Dubnau andDavidoff-Abelson, 1971, J. Mol. Biol. 56: 209-221), electroporation(see, e.g., Shigekawa and Dower, 1988, Biotechniques 6: 742-751), orconjugation (see, e.g., Koehler and Thorne, 1987, J. Bacteriol. 169:5271-5278). The introduction of DNA into an E. coli cell may be effectedby protoplast transformation (see, e.g., Hanahan, 1983, J. Mol. Biol.166: 557-580) or electroporation (see, e.g., Dower et al., 1988, NucleicAcids Res. 16: 6127-6145). The introduction of DNA into a Streptomycescell may be effected by protoplast transformation, electroporation (see,e.g., Gong et al., 2004, Folia Microbiol. (Praha) 49: 399-405),conjugation (see, e.g., Mazodier et al., 1989, J. Bacteriol. 171:3583-3585), or transduction (see, e.g., Burke et al., 2001, Proc. Natl.Acad. Sci. USA 98: 6289-6294). The introduction of DNA into aPseudomonas cell may be effected by electroporation (see, e.g., Choi etal., 2006, J. Microbiol. Methods 64: 391-397) or conjugation (see, e.g.,Pinedo and Smets, 2005, Appl. Environ. Microbiol. 71: 51-57). Theintroduction of DNA into a Streptococcus cell may be effected by naturalcompetence (see, e.g., Perry and Kuramitsu, 1981, Infect. Immun. 32:1295-1297), protoplast transformation (see, e.g., Catt and Jollick,1991, Microbios 68: 189-207), electroporation (see, e.g., Buckley etal., 1999, Appl. Environ. Microbiol. 65: 3800-3804), or conjugation(see, e.g., Clewell, 1981, Microbiol. Rev. 45: 409-436). However, anymethod known in the art for introducing DNA into a host cell can beused.

The host cell may also be a eukaryote, such as a mammalian, insect,plant, or fungal cell.

The host cell may be a fungal cell. “Fungi” as used herein includes thephyla Ascomycota, Basidiomycota, Chytridiomycota, and Zygomycota as wellas the Oomycota and all mitosporic fungi (as defined by Hawksworth etal., In, Ainsworth and Bisby's Dictionary of The Fungi, 8th edition,1995, CAB International, University Press, Cambridge, UK).

The fungal host cell may be a yeast cell. “Yeast” as used hereinincludes ascosporogenous yeast (Endomycetales), basidiosporogenousyeast, and yeast belonging to the Fungi Imperfecti (Blastomycetes).Since the classification of yeast may change in the future, for thepurposes of this invention, yeast shall be defined as described inBiology and Activities of Yeast (Skinner, Passmore, and Davenport,editors, Soc. App. Bacteriol. Symposium Series No. 9, 1980).

The yeast host cell may be a Candida, Hansenula, Kluyveromyces, Pichia,Saccharomyces, Schizosaccharomyces, or Yarrowia cell, such as aKluyveromyces lactis, Saccharomyces carlsbergensis, Saccharomycescerevisiae, Saccharomyces diastaticus, Saccharomyces douglasii,Saccharomyces kluyveri, Saccharomyces norbensis, Saccharomycesoviformis, or Yarrowia lipolytica cell.

The fungal host cell may be a filamentous fungal cell. “Filamentousfungi” include all filamentous forms of the subdivision Eumycota andOomycota (as defined by Hawksworth et al., 1995, supra). The filamentousfungi are generally characterized by a mycelial wall composed of chitin,cellulose, glucan, chitosan, mannan, and other complex polysaccharides.Vegetative growth is by hyphal elongation and carbon catabolism isobligately aerobic. In contrast, vegetative growth by yeasts such asSaccharomyces cerevisiae is by budding of a unicellular thallus andcarbon catabolism may be fermentative.

The filamentous fungal host cell may be an Acremonium, Aspergillus,Aureobasidium, Bjerkandera, Ceriporiopsis, Chrysosporium, Coprinus,Coriolus, Cryptococcus, Filibasidium, Fusarium, Humicola, Magnaporthe,Mucor, Myceliophthora, Neocallimastix, Neurospora, Paecilomyces,Penicillium, Phanerochaete, Phlebia, Piromyces, Pleurotus,Schizophyllum, Talaromyces, Thermoascus, Thielavia, Tolypocladium,Trametes, or Trichoderma cell.

For example, the filamentous fungal host cell may be an Aspergillusawamori, Aspergillus foetidus, Aspergillus fumigatus, Aspergillusjaponicus, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae,Bjerkandera adusta, Ceriporiopsis aneirina, Ceriporiopsis caregiea,Ceriporiopsis gilvescens, Ceriporiopsis pannocinta, Ceriporiopsisrivulosa, Ceriporiopsis subrufa, Ceriporiopsis subvermispora,Chrysosporium inops, Chrysosporium keratinophilum, Chrysosporiumlucknowense, Chrysosporium merdarium, Chrysosporium pannicola,Chrysosporium queenslandicum, Chrysosporium tropicum, Chrysosporiumzonatum, Coprinus cinereus, Coriolus hirsutus, Fusarium bactridioides,Fusarium cerealis, Fusarium crookwellense, Fusarium culmorum, Fusariumgraminearum, Fusarium graminum, Fusarium heterosporum, Fusarium negundi,Fusarium oxysporum, Fusarium reticulatum, Fusarium roseum, Fusariumsambucinum, Fusarium sarcochroum, Fusarium sporotrichioides, Fusariumsulphureum, Fusarium torulosum, Fusarium trichothecioides, Fusariumvenenatum, Humicola insolens, Humicola lanuginosa, Mucor miehei,Myceliophthora thermophila, Neurospora crassa, Penicillium purpurogenum,Phanerochaete chrysosporium, Phlebia radiata, Pleurotus eryngii,Thielavia terrestris, Trametes villosa, Trametes versicolor, Trichodermaharzianum, Trichoderma koningii, Trichoderma longibrachiatum,Trichoderma reesei, or Trichoderma viride cell.

Fungal cells may be transformed by a process involving protoplastformation, transformation of the protoplasts, and regeneration of thecell wall in a manner known per se. Suitable procedures fortransformation of Aspergillus and Trichoderma host cells are describedin EP 238023, Yelton et al., 1984, Proc. Natl. Acad. Sci. USA 81:1470-1474, and Christensen et al., 1988, Bio/Technology 6: 1419-1422.Suitable methods for transforming Fusarium species are described byMalardier et al., 1989, Gene 78: 147-156, and WO 96/00787. Yeast may betransformed using the procedures described by Becker and Guarente, InAbelson, J. N. and Simon, M. I., editors, Guide to Yeast Genetics andMolecular Biology, Methods in Enzymology, Volume 194, pp 182-187,Academic Press, Inc., New York; Ito et al., 1983, J. Bacteriol. 153:163; and Hinnen et al., 1978, Proc. Natl. Acad. Sci. USA 75: 1920.

Methods of Production

The present invention also relates to methods of producing a polypeptideof the present invention, comprising (a) cultivating a cell, which inits wild-type form produces the polypeptide, under conditions conducivefor production of the polypeptide; and optionally, (b) recovering thepolypeptide. In one aspect, the cell is an Aspergillus cell. In anotheraspect, the cell is an Aspergillus oryzae cell.

The present invention also relates to methods of producing a polypeptideof the present invention, comprising (a) cultivating a recombinant hostcell of the present invention under conditions conducive for productionof the polypeptide; and optionally, (b) recovering the polypeptide.

The host cells are cultivated in a nutrient medium suitable forproduction of the polypeptide using methods known in the art. Forexample, the cells may be cultivated by shake flask cultivation, orsmall-scale or large-scale fermentation (including continuous, batch,fed-batch, or solid state fermentations) in laboratory or industrialfermentors in a suitable medium and under conditions allowing thepolypeptide to be expressed and/or isolated. The cultivation takes placein a suitable nutrient medium comprising carbon and nitrogen sources andinorganic salts, using procedures known in the art. Suitable media areavailable from commercial suppliers or may be prepared according topublished compositions (e.g., in catalogues of the American Type CultureCollection). If the polypeptide is secreted into the nutrient medium,the polypeptide can be recovered directly from the medium. If thepolypeptide is not secreted, it can be recovered from cell lysates.

The polypeptide may be detected using methods known in the art that arespecific for the polypeptides. These detection methods include, but arenot limited to, use of specific antibodies, formation of an enzymeproduct, or disappearance of an enzyme substrate. For example, an enzymeassay may be used to determine the activity of the polypeptide.

The polypeptide may be recovered using methods known in the art. Forexample, the polypeptide may be recovered from the nutrient medium byconventional procedures including, but not limited to, collection,centrifugation, filtration, extraction, spray-drying, evaporation, orprecipitation. In one aspect, a fermentation broth comprising thepolypeptide is recovered.

The polypeptide may be purified by a variety of procedures known in theart including, but not limited to, chromatography (e.g., ion exchange,affinity, hydrophobic, chromatofocusing, and size exclusion),electrophoretic procedures (e.g., preparative isoelectric focusing),differential solubility (e.g., ammonium sulfate precipitation),SDS-PAGE, or extraction (see, e.g., Protein Purification, Janson andRyden, editors, VCH Publishers, New York, 1989) to obtain substantiallypure polypeptides.

In an alternative aspect, the polypeptide is not recovered, but rather ahost cell of the present invention expressing the polypeptide is used asa source of the polypeptide.

In one embodiment, the invention further comprises producing thepolypeptide by cultivating the recombinant host cell further comprisinga polynucleotide encoding a second polypeptide of interest; preferablyan enzyme of interest; more preferably a secreted enzyme of interest;even more preferably a hydrolase, isomerase, ligase, lyase,oxidoreductase, or a transferase; and most preferably the secretedenzyme is an alpha-galactosidase, alpha-glucosidase, aminopeptidase,amylase, asparaginase, beta-galactosidase, beta-glucosidase,beta-xylosidase, carbohydrase, carboxypeptidase, catalase,cellobiohydrolase, cellulase, chitinase, cutinase, cyclodextringlycosyltransferase, deoxyribonuclease, endoglucanase, esterase, greenfluorescent protein, glucano-transferase, glucoamylase, invertase,laccase, lipase, mannosidase, mutanase, oxidase, pectinolytic enzyme,peroxidase, phytase, polyphenoloxidase, proteolytic enzyme,ribonuclease, transglutaminase, or a xylanase.

In one embodiment, the second polypeptide of interest is heterologous orhomologous to the host cell.

In one embodiment, the recombinant host cell is a fungal host cell;preferably a filamentous fungal host cell; more preferably anAcremonium, Aspergillus, Aureobasidium, Bjerkandera, Ceriporiopsis,Chrysosporium, Coprinus, Coriolus, Cryptococcus, Filibasidium, Fusarium,Humicola, Magnaporthe, Mucor, Myceliophthora, Neocallimastix,Neurospora, Paecilomyces, Penicillium, Phanerochaete, Phlebia,Piromyces, Pleurotus, Schizophyllum, Talaromyces, Thermoascus,Thielavia, Tolypocladium, Trametes, or Trichoderma cell; most preferablyan Aspergillus awamori, Aspergillus foetidus, Aspergillus fumigatus,Aspergillus japonicus, Aspergillus nidulans, Aspergillus niger,Aspergillus oryzae, Bjerkandera adusta, Ceriporiopsis aneirina,Ceriporiopsis caregiea, Ceriporiopsis gilvescens, Ceriporiopsispannocinta, Ceriporiopsis rivulosa, Ceriporiopsis subrufa, Ceriporiopsissubvermispora, Chrysosporium inops, Chrysosporium keratinophilum,Chrysosporium lucknowense, Chrysosporium merdarium, Chrysosporiumpannicola, Chrysosporium queenslandicum, Chrysosporium tropicum,Chrysosporium zonatum, Coprinus cinereus, Coriolus hirsutus, Fusariumbactridioides, Fusarium cerealis, Fusarium crookwellense, Fusariumculmorum, Fusarium graminearum, Fusarium graminum, Fusariumheterosporum, Fusarium negundi, Fusarium oxysporum, Fusariumreticulatum, Fusarium roseum, Fusarium sambucinum, Fusarium sarcochroum,Fusarium sporotrichioides, Fusarium sulphureum, Fusarium torulosum,Fusarium trichothecioides, Fusarium venenatum, Humicola insolens,Humicola lanuginosa, Mucor miehei, Myceliophthora thermophila,Neurospora crassa, Penicillium purpurogenum, Phanerochaetechrysosporium, Phlebia radiata, Pleurotus eryngii, Thielavia terrestris,Trametes villosa, Trametes versicolor, Trichoderma harzianum,Trichoderma koningii, Trichoderma longibrachiatum, Trichoderma reesei,or Trichoderma viride cell.

In one embodiment, the recombinant host cell is a bacterial host cell;preferably a prokaryotic host cell; more preferably a Gram-positive hostcell; even more preferably a Bacillus, Clostridium, Enterococcus,Geobacillus, Lactobacillus, Lactococcus, Oceanobacillus, Staphylococcus,Streptococcus, or Streptomyces host cell; and most preferably a Bacillusalkalophilus, Bacillus amyloliquefaciens, Bacillus brevis, Bacilluscirculans, Bacillus clausii, Bacillus coagulans, Bacillus firmus,Bacillus lautus, Bacillus lentus, Bacillus licheniformis, Bacillusmegaterium, Bacillus pumilus, Bacillus stearothermophilus, Bacillussubtilis, and Bacillus thuringiensis host cell.

In one embodiment, a method of producing the second polypeptide ofinterest comprises cultivating the host cell under conditions conducivefor production of the second polypeptide of interest.

In one embodiment, the method further comprises recovering the secondpolypeptide of interest.

Compositions

The present invention further concerns a detergent compositioncomprising at least one DNase variant according to the invention andpreferably a detergent adjunct ingredient. The detergent composition maybe used for preventing, reducing or removing biofilm from an item, forpreventing, reducing or removing the stickiness of an item, forpretreating stains on the item, for preventing, reducing or removingredeposition of soil during a wash cycle, for reducing or removingadherence of soil to an item, for maintaining or improving the whitenessof an item and for preventing, reducing or removing malodor from anitem, such as E-2-nonenal as described in Assay II. The detergentcompositions comprising the polypeptides of the present inventionovercomes the problems of the prior art. The DNase variants of theinvention are useful in powder and liquid detergent.

One embodiment of the invention relates to a detergent compositioncomprising a DNase variant which compared to SEQ ID NO: 1 comprises analteration at one or more positions selected from the group consistingof: 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19,20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37,38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 53, 54, 55, 56,57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74,75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92,93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108,109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122,123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136,137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150,151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164,165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177, 178,179, 180, 181, 182, 183, 185, 186, 187, 189, 190, 191, 192, 193, 194,195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207, 208,209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220 and 221 and adetergent adjunct.

In one embodiment of the invention, the detergent composition comprisesa DNase variant which compared to SEQ ID NO: 1 comprises one or morealterations selected from the group consisting of: V1*, V1A, V1D, V1E,V1F, V1G, V1H, V1I, V1K, V1L, V1M, V1N, V1P, V1Q, V1R, V1S, V1T, V1W,V1Y, P2*, P2A, P2D, P2E, P2F, P2G, P2H, P2I, P2K, P2L, P2M, P2N, P2Q,P2R, P2S, P2T, P2V, P2W, P2Y, V3*, V3A, V3C, V3D, V3E, V3F, V3G, V3H,V3I, V3K, V3L, V3M, V3N, V3P, V3R, V3S, V3T, V3W, V3Y, N4*, N4A, N4D,N4E, N4F, N4G, N4H, N4I, N4K, N4L, N4M, N4P, N4Q, N4R, N4S, N4T, N4V,N4W, N4Y, P5*, P5A, P5D, P5E, P5F, P5G, P5H, P5I, P5K, P5L, P5M, P5N,P5Q, P5R, P5S, P5T, P5V, P5W, P5Y, E6*, E6A, E6D, E6F, E6G, E6H, E6I,E6K, E6L, E6M, E6N, E6P, E6Q, E6R, E6S, E6T, E6V, E6W, E6Y, P7*, P7A,P7D, P7E, P7F, P7G, P7H, P7I, P7K, P7L, P7M, P7N, P7Q, P7R, P7S, P7T,P7V, P7W, P7Y, D8*, D8A, D8E, D8F, D8G, D8H, D8I, D8K, D8L, D8M, D8N,D8P, D8Q, D8R, D8S, D8T, D8V, D8W, D8Y, A9*, A9D, A9E, A9F, A9G, A9H,A9I, A9K, A9L, A9M, A9N, A9P, A9Q, A9R, A9S, A9T, A9V, A9W, A9Y, T10*,T10A, T10D, T10E, T10F, T10G, T10H, T10I, T10K, T10L, T10M, T10N, T10P,T10Q, T10R, T10S, T10V, T10W, T10Y, S11*, S11A, S11D, S11E, S11F, S11G,S11H, S11I, S11K, S11L, S11M, S11N, S11P, S11Q, S11R, S11T, S11V, S11W,S11Y, V12*, V12A, V12D, V12E, V12F, V12G, V12H, V12I, V12K, V12L, V12M,V12N, V12P, V12Q, V12R, V12S, V12T, V12W, V12Y, E13*, E13A, E13D, E13F,E13G, E13H, E13I, E13K, E13L, E13M, E13N, E13P, E13Q, E13R, E13S, E13T,E13V, E13W, E13Y, N14*, N14A, N14D, N14E, N14F, N14G, N14H, N14I, N14K,N14L, N14M, N14P, N14Q, N14R, N14S, N14T, N14V, N14W, N14Y, V15*, V15A,V15D, V15E, V15F, V15G, V15H, V15I, V15K, V15L, V15M, V15N, V15P, V15Q,V15R, V15S, V15T, V15W, V15Y, A16*, A16D, A16E, A16F, A16G, A16H, A16I,A16K, A16L, A16M, A16N, A16P, A16Q, A16R, A16S, A16T, A16V, A16W, A16Y,L17*, L17A, L17D, L17E, L17F, L17G, L17H, L17I, L17K, L17M, L17N, L17P,L17Q, L17R, L17S, L17T, L17V, L17W, L17Y, K18*, K18A, K18D, K18E, K18F,K18G, K18H, K18I, K18L, K18M, K18N, K18P, K18Q, K18R, K18S, K18T, K18V,K18W, K18Y, T19*, T19A, T19D, T19E, T19F, T19G, T19H, T19I, T19K, T19L,T19M, T19N, T19P, T19Q, T19R, T19S, T19V, T19W, T19Y, G20*, G20A, G20D,G20E, G20F, G20H, G20I, G20K, G20L, G20M, G20N, G20P, G20Q, G20R, G20S,G20T, G20V, G20W, G20Y, S21*, S21A, S21D, S21E, S21F, S21G, S21H, S21I,S21K, S21L, S21M, S21N, S21P, S21Q, S21R, S21T, S21V, S21W, S21Y, G22*,G22A, G22D, G22E, G22F, G22H, G22I, G22K, G22L, G22M, G22N, G22P, G22Q,G22R, G22S, G22T, G22V, G22W, G22Y, D23*, D23A, D23E, D23F, D23G, D23H,D23I, D23K, D23L, D23M, D23N, D23P, D23Q, D23R, D23S, D23T, D23V, D23W,D23Y, S24*, S24A, S24D, S24E, S24F, S24G, S24H, S24I, S24K, S24L, S24M,S24N, S24P, S24Q, S24R, S24T, S24V, S24W, S24Y, Q25*, Q25A, Q25D, Q25E,Q25F, Q25G, Q25H, Q25I, Q25K, Q25L, Q25M, Q25N, Q25P, Q25R, Q25S, Q25T,Q25V, Q25W, Q25Y, S26*, S26A, S26D, S26E, S26F, S26G, S26H, S26I, S26K,S26L, S26M, S26N, S26P, S26Q, S26R, S26T, S26V, S26W, S26Y, D27*, D27A,D27E, D27F, D27G, D27H, D27I, D27K, D27L, D27M, D27N, D27P, D27Q, D27R,D27S, D27T, D27V, D27W, D27Y, P28*, P28A, P28D, P28E, P28F, P28G, P28H,P28I, P28K, P28L, P28M, P28N, P28Q, P28R, P28S, P28T, P28V, P28W, P28Y,I29*, I29A, I29D, I29E, I29F, I29G, I29H, I29K, I29L, I29M, I29N, I29P,I29Q, I29R, I29S, I29T, I29V, I29W, I29Y, K30*, K30A, K30D, K30E, K30F,K30G, K30H, K30I, K30L, K30M, K30N, K30P, K30Q, K30R, K30S, K30T, K30V,K30W, K30Y, A31*, A31D, A31E, A31F, A31G, A31H, A31I, A31K, A31L, A31M,A31N, A31P, A31Q, A31R, A31S, A31T, A31V, A31W, A31Y, D32*, D32A, D32E,D32F, D32G, D32H, D32I, D32K, D32L, D32M, D32N, D32P, D32Q, D32R, D32S,D32T, D32V, D32W, D32Y, L33*, L33A, L33D, L33E, L33F, L33G, L33H, L33I,L33K, L33M, L33N, L33P, L33Q, L33R, L33S, L33T, L33V, L33W, L33Y, E34*,E34A, E34D, E34F, E34G, E34H, E34I, E34K, E34L, E34M, E34N, E34P, E34Q,E34R, E34S, E34T, E34V, E34W, E34Y, V35*, V35A, V35D, V35E, V35F, V35G,V35H, V35I, V35K, V35L, V35M, V35N, V35P, V35Q, V35R, V35S, V35T, V35W,V35Y, K36*, K36A, K36D, K36E, K36F, K36G, K36H, K36I, K36L, K36M, K36N,K36P, K36Q, K36R, K36S, K36T, K36V, K36W, K36Y, G37*, G37A, G37D, G37E,G37F, G37H, G37I, G37K, G37L, G37M, G37N, G37P, G37Q, G37R, G37S, G37T,G37V, G37W, G37Y, Q38*, Q38A, Q38D, Q38E, Q38F, Q38G, Q38H, Q38I, Q38K,Q38L, Q38M, Q38N, Q38P, Q38R, Q38S, Q38T, Q38V, Q38W, Q38Y, S39*, S39A,S39D, S39E, S39F, S39G, S39H, S39I, S39K, S39L, S39M, S39N, S39P, S39Q,S39R, S39T, S39V, S39W, S39Y, A40*, A40D, A40E, A40F, A40G, A40H, A40I,A40K, A40L, A40M, A40N, A40P, A40Q, A40R, A40S, A40T, A40V, A40W, A40Y,L41*, L41A, L41D, L41E, L41F, L41G, L41H, L41I, L41K, L41M, L41N, L41P,L41Q, L41R, L41S, L41T, L41V, L41W, L41Y, P42*, P42A, P42D, P42E, P42F,P42G, P42H, P42I, P42K, P42L, P42M, P42N, P42Q, P42R, P42S, P42T, P42V,P42W, P42Y, F43*, F43A, F43D, F43E, F43G, F43H, F43I, F43K, F43L, F43M,F43N, F43P, F43Q, F43R, F43S, F43T, F43V, F43W, F43Y, D44*, D44A, D44E,D44F, D44G, D44H, D44I, D44K, D44L, D44M, D44N, D44P, D44Q, D44R, D44S,D44T, D44V, D44W, D44Y, V45*, V45A, V45D, V45E, V45F, V45G, V45H, V45I,V45K, V45L, V45M, V45N, V45P, V45Q, V45R, V45S, V45T, V45W, V45Y, D46*,D46A, D46E, D46F, D46G, D46H, D46I, D46K, D46L, D46M, D46N, D46P, D46Q,D46R, D46S, D46T, D46V, D46W, D46Y, C47A, C47D, C47E, C47F, C47G, C47H,C47I, C47K, C47L, C47M, C47N, C47P, C47Q, C47R, C47S, C47T, C47V, C47W,C47Y, W48*, W48A, W48D, W48E, W48F, W48G, W48H, W48I, W48K, W48L, W48M,W48N, W48P, W48Q, W48R, W48S, W48T, W48V, W48Y, A49*, A49D, A49E, A49F,A49G, A49H, A49I, A49K, A49L, A49M, A49N, A49P, A49Q, A49R, A49S, A49T,A49V, A49W, A49Y, 150*, 150A, I50D, I50E, I50F, I50G, I50H, I50K, I50L,I50M, I50N, I50P, I50Q, I50R, I50S, I50T, I50V, I50W, 150Y, L51*, L51A,L51D, L51E, L51F, L51G, L51H, L51I, L51K, L51M, L51N, L51P, L51Q, L51R,L51S, L51T, L51V, L51W, L51Y, K53*, K53A, K53D, K53E, K53F, K53G, K53H,K53I, K53L, K53M, K53N, K53P, K53Q, K53R, K53S, K53T, K53V, K53W, K53Y,G54*, G54A, G54D, G54E, G54F, G54H, G54I, G54K, G54L, G54M, G54N, G54P,G54Q, G54R, G54S, G54T, G54V, G54W, G54Y, A55*, A55D, A55E, A55F, A55G,A55H, A55I, A55K, A55L, A55M, A55N, A55P, A55Q, A55R, A55S, A55T, A55V,A55W, A55Y, P56*, P56A, P56D, P56E, P56F, P56G, P56H, P56I, P56K, P56L,P56M, P56N, P56Q, P56R, P56S, P56T, P56V, P56W, P56Y, N57*, N57A, N57D,N57E, N57F, N57G, N57H, N57I, N57K, N57L, N57M, N57P, N57Q, N57R, N57S,N57T, N57V, N57W, N57Y, V58*, V58A, V58D, V58E, V58F, V58G, V58H, V58I,V58K, V58L, V58M, V58N, V58P, V58Q, V58R, V58S, V58T, V58W, V58YL59*,L59A, L59D, L59E, L59F, L59G, L59H, L59I, L59K, L59M, L59N, L59P, L59Q,L59R, L59S, L59T, L59V, L59W, L59Y, Q60*, Q60A, Q60D, Q60E, Q60F, Q60G,Q60H, Q60I, Q60K, Q60L, Q60M, Q60N, Q60P, Q60R, Q60S, Q60T, Q60V, Q60W,Q60Y, R61*, R61A, R61D, R61E, R61F, R61G, R61H, R61I, R61K, R61L, R61M,R61N, R61P, R61Q, R61S, R61T, R61V, R61W, R61Y, V62*, V62A, V62D, V62E,V62F, V62G, V62H, V62I, V62K, V62L, V62M, V62N, V62P, V62Q, V62R, V62S,V62T, V62W, V62Y, N63*, N63A, N63D, N63E, N63F, N63G, N63H, N63I, N63K,N63L, N63M, N63P, N63Q, N63R, N63S, N63T, N63V, N63W, N63Y, E64*, E64A,E64D, E64F, E64G, E64H, E64I, E64K, E64L, E64M, E64N, E64P, E64Q, E64R,E64S, E64T, E64V, E64W, E64Y, K65*, K65A, K65D, K65E, K65F, K65G, K65H,K65I, K65L, K65M, K65N, K65P, K65Q, K65R, K65S, K65T, K65V, K65W, K65Y,T66*, T66A, T66D, T66E, T66F, T66G, T66H, T66I, T66K, T66L, T66M, T66N,T66P, T66Q, T66R, T66S, T66V, T66W, T66Y, K67*, K67A, K67D, K67E, K67F,K67G, K67H, K67I, K67L, K67M, K67N, K67P, K67Q, K67R, K67S, K67T, K67V,K67W, K67Y, N68*, N68A, N68D, N68E, N68F, N68G, N68H, N68I, N68K, N68L,N68M, N68P, N68Q, N68R, N68S, N68T, N68V, N68W, N68Y, S69*, S69A, S69D,S69E, S69F, S69G, S69H, S69I, S69K, S69L, S69M, S69N, S69P, S69Q, S69R,S69T, S69V, S69W, S69Y, N70*, N70A, N70D, N70E, N70F, N70G, N70H, N70I,N70K, N70L, N70M, N70P, N70Q, N70R, N70S, N70T, N70V, N70W, N70Y, R71*,R71A, R71D, R71E, R71F, R71G, R71H, R71I, R71K, R71L, R71M, R71N, R71P,R71Q, R71S, R71T, R71V, R71W, R71Y, D72*, D72A, D72E, D72F, D72G, D72H,D72I, D72K, D72L, D72M, D72N, D72P, D72Q, D72R, D72S, D72T, D72V, D72W,D72Y, R73*, R73A, R73D, R73E, R73F, R73G, R73H, R73I, R73K, R73L, R73M,R73N, R73P, R73Q, R73S, R73T, R73V, R73W, R73Y, S74*, S74A, S74D, S74E,S74F, S74G, S74H, S74I, S74K, S74L, S74M, S74N, S74P, S74Q, S74R, S74T,S74V, S74W, S74Y, G75*, G75A, G75D, G75E, G75F, G75H, G75I, G75K, G75L,G75M, G75N, G75P, G75Q, G75R, G75S, G75T, G75V, G75W, G75Y, A76*, A76D,A76E, A76F, A76G, A76H, A76I, A76K, A76L, A76M, A76N, A76P, A76Q, A76R,A76S, A76T, A76V, A76W, A76Y, N77*, N77A, N77D, N77E, N77F, N77G, N77H,N77I, N77K, N77L, N77M, N77P, N77Q, N77R, N77S, N77T, N77V, N77W, N77Y,K78*, K78A, K78D, K78E, K78F, K78G, K78H, K78I, K78L, K78M, K78N, K78P,K78Q, K78R, K78S, K78T, K78V, K78W, K78Y, G79*, G79A, G79D, G79E, G79F,G79H, G79I, G79K, G79L, G79M, G79N, G79P, G79Q, G79R, G79S, G79T, G79V,G79W, G79Y, P80*, P80A, P80D, P80E, P80F, P80G, P80H, P80I, P80K, P80L,P80M, P80N, P80Q, P80R, P80S, P80T, P80V, P80W, P80Y, F81*, F81A, F81D,F81E, F81G, F81H, F81I, F81K, F81L, F81M, F81N, F81P, F81Q, F81R, F81S,F81T, F81V, F81W, F81Y, K82*, K82A, K82D, K82E, K82F, K82G, K82H, K82I,K82L, K82M, K82N, K82P, K82Q, K82R, K82S, K82T, K82V, K82W, K82Y, D83*,D83A, D83E, D83F, D83G, D83H, D83I, D83K, D83L, D83M, D83N, D83P, D83Q,D83R, D83S, D83T, D83V, D83W, D83Y, P84*, P84A, P84D, P84E, P84F, P84G,P84H, P84I, P84K, P84L, P84M, P84N, P84Q, P84R, P84S, P84T, P84V, P84W,P84Y, Q85*, Q85A, Q85D, Q85E, Q85F, Q85G, Q85H, Q85I, Q85K, Q85L, Q85M,Q85N, Q85P, Q85R, Q85S, Q85T, Q85V, Q85W, Q85Y, K86*, K86A, K86D, K86E,K86F, K86G, K86H, K86I, K86L, K86M, K86N, K86P, K86Q, K86R, K86S, K86T,K86V, K86W, K86Y, W87*, W87A, W87D, W87E, W87F, W87G, W87H, W87I, W87K,W87L, W87M, W87N, W87P, W87Q, W87R, W87S, W87T, W87V, W87Y, G88*, G88A,G88D, G88E, G88F, G88H, G88I, G88K, G88L, G88M, G88N, G88P, G88Q, G88R,G88S, G88T, G88V, G88W, G88Y, I89*, I89A, I89D, I89E, I89F, I89G, I89H,I89K, I89L, I89M, I89N, I89P, I89Q, I89R, I89S, I89T, I89V, I89W, I89Y,K90*, K90A, K90D, K90E, K90F, K90G, K90H, K90I, K90L, K90M, K90N, K90P,K90Q, K90R, K90S, K90T, K90V, K90W, K90Y, A91*, A91D, A91E, A91F, A91G,A91H, A91I, A91K, A91L, A91M, A91N, A91P, A91Q, A91R, A91S, A91T, A91V,A91W, A91Y, L92*, L92A, L92D, L92E, L92F, L92G, L92H, L92I, L92K, L92M,L92N, L92P, L92Q, L92R, L92S, L92T, L92V, L92W, L92Y, P93*, P93A, P93D,P93E, P93F, P93G, P93H, P93I, P93K, P93L, P93M, P93N, P93Q, P93R, P93S,P93T, P93V, P93W, P93Y, P94*, P94A, P94D, P94E, P94F, P94G, P94H, P94I,P94K, P94L, P94M, P94N, P94Q, P94R, P94S, P94T, P94V, P94W, P94Y, K95*,K95A, K95D, K95E, K95F, K95G, K95H, K95I, K95L, K95M, K95N, K95P, K95Q,K95R, K95S, K95T, K95V, K95W, K95Y, N96*, N96A, N96D, N96E, N96F, N96G,N96H, N96I, N96K, N96L, N96M, N96P, N96Q, N96R, N96S, N96T, N96V, N96W,N96Y, P97*, P97A, P97D, P97E, P97F, P97G, P97H, P97I, P97K, P97L, P97M,P97N, P97Q, P97R, P97S, P97T, P97V, P97W, P97Y, S98*, S98A, S98D, S98E,S98F, S98G, S98H, S98I, S98K, S98L, S98M, S98N, S98P, S98Q, S98R, S98T,S98V, S98W, S98Y, W99*, W99A, W99D, W99E, W99F, W99G, W99H, W99I, W99K,W99L, W99M, W99N, W99P, W99Q, W99R, W99S, W99T, W99V, W99Y, S100*,S100A, S100D, S100E, S100F, S100G, S100H, S100I, S100K, S100L, S100M,S100N, S100P, S100Q, S100R, S100T, S100V, S100W, S100Y, A101*, A101D,A101E, A101F, A101G, A101H, A101I, A101K, A101L, A101M, A101N, A101P,A101Q, A101R, A101S, A101T, A101V, A101W, A101Y, Q102*, Q102A, Q102D,Q102E, Q102F, Q102G, Q102H, Q102I, Q102K, Q102L, Q102M, Q102N, Q102P,Q102R, Q102S, Q102T, Q102V, Q102W, Q102Y, D103*, D103A, D103E, D103F,D103G, D103H, D103I, D103K, D103L, D103M, D103N, D103P, D103Q, D103R,D103S, D103T, D103V, D103W, D103Y, F104*, F104A, F104D, F104E, F104G,F104H, F104I, F104K, F104L, F104M, F104N, F104P, F104Q, F104R, F104S,F104T, F104V, F104W, F104Y, K105*, K105A, K105D, K105E, K105F, K105G,K105H, K105I, K105L, K105M, K105N, K105P, K105Q, K105R, K105S, K105T,K105V, K105W, K105Y, S106*, S106A, S106D, S106E, S106F, S106G, S106H,S106I, S106K, S106L, S106M, S106N, S106P, S106Q, S106R, S106T, S106V,S106W, S106Y, P107*, P107A, P107D, P107E, P107F, P107G, P107H, P107I,P107K, P107L, P107M, P107N, P107Q, P107R, P107S, P107T, P107V, P107W,P107Y, E108*, E108A, E108D, E108F, E108G, E108H, E108I, E108K, E108L,E108M, E108N, E108P, E108Q, E108R, E108S, E108T, E108V, E108W, E108Y,E109*, E109A, E109D, E109F, E109G, E109H, E109I, E109K, E109L, E109M,E109N, E109P, E109Q, E109R, E109S, E109T, E109V, E109W, E109Y, Y110*,Y110A, Y110D, Y110E, Y110F, Y110G, Y110H, Y110I, Y110K, Y110L, Y110M,Y110N, Y110P, Y110Q, Y110R, Y110S, Y110T, Y110V, Y110W, A111*, A111D,A111E, A111F, A111G, A111H, A111I, A111K, A111L, A111M, A111N, A111P,A111Q, A111R, A111S, A111T, A111V, A111W, A111Y, F112*, F112A, F112D,F112E, F112G, F112H, F112I, F112K, F112L, F112M, F112N, F112P, F112Q,F112R, F112S, F112T, F112V, F112W, F112Y, A113*, A113D, A113E, A113F,A113G, A113H, A113I, A113K, A113L, A113M, A113N, A113P, A113Q, A113R,A113S, A113T, A113V, A113W, A113Y, S114*, S114A, S114D, S114E, S114F,S114G, S114H, S114I, S114K, S114L, S114M, S114N, S114P, S114Q, S114R,S114T, S114V, S114W, S114Y, S115*, S115A, S115D, S115E, S115F, S115G,S115H, S115I, S115K, S115L, S115M, S115N, S115P, S115Q, S115R, S115T,S115V, S115W, S115Y, L116*, L116A, L116D, L116E, L116F, L116G, L116H,L116I, L116K, L116M, L116N, L116P, L116Q, L116R, L116S, L116T, L116V,L116W, L116Y, Q117*, Q117A, Q117D, Q117E, Q117F, Q117G, Q117H, Q117I,Q117K, Q117L, Q117M, Q117N, Q117P, Q117R, Q117S, Q117T, Q117V, Q117W,Q117Y, G118*, G118A, G118D, G118E, G118F, G118H, G118I, G118K, G118L,G118M, G118N, G118P, G118Q, G118R, G118S, G118T, G118V, G118W, G118Y,G119*, G119A, G119D, G119E, G119F, G119H, G119I, G119K, G119L, G119M,G119N, G119P, G119Q, G119R, G119S, G119T, G119V, G119W, G119Y, T120*,T120A, T120D, T120E, T120F, T120G, T120H, T120I, T120K, T120L, T120M,T120N, T120P, T120Q, T120R, T120S, T120V, T120W, T120Y, N121*, N121A,N121D, N121E, N121F, N121G, N121H, N121I, N121K, N121L, N121M, N121P,N121Q, N121R, N121S, N121T, N121V, N121W, N121Y, A122*, A122D, A122E,A122F, A122G, A122H, A122I, A122K, A122L, A122M, A122N, A122P, A122Q,A122R, A122S, A122T, A122V, A122W, A122Y, I123*, I123A, I123D, I123E,I123F, I123G, I123H, I123K, I123L, I123M, I123N, I123P, I123Q, I123R,I123S, I123T, I123V, I123W, I123Y, L124*, L124A, L124D, L124E, L124F,L124G, L124H, L124I, L124K, L124M, L124N, L124P, L124Q, L124R, L124S,L124T, L124V, L124W, L124Y, A125*, A125D, A125E, A125F, A125G, A125H,A125I, A125K, A125L, A125M, A125N, A125P, A125Q, A125R, A125S, A125T,A125V, A125W, A125Y, P126*, P126A, P126D, P126E, P126F, P126G, P126H,P126I, P126K, P126L, P126M, P126N, P126Q, P126R, P126S, P126T, P126V,P126W, P126Y, V127*, V127A, V127D, V127E, V127F, V127G, V127H, V127I,V127K, V127L, V127M, V127N, V127P, V127Q, V127R, V127S, V127T, V127W,V127Y, N128*, N128A, N128D, N128E, N128F, N128G, N128H, N128I, N128K,N128L, N128M, N128P, N128Q, N128R, N128S, N128T, N128V, N128W, N128Y,L129*, L129A, L129D, L129E, L129F, L129G, L129H, L129I, L129K, L129M,L129N, L129P, L129Q, L129R, L129S, L129T, L129V, L129W, L129Y, A130*,A130D, A130E, A130F, A130G, A130H, A130I, A130K, A130L, A130M, A130N,A130P, A130Q, A130R, A130S, A130T, A130V, A130W, A130Y, S131*, S131A,S131D, S131E, S131F, S131G, S131H, S131I, S131K, S131L, S131M, S131N,S131P, S131Q, S131R, S131T, S131V, S131W, S131Y, Q132*, Q132A, Q132D,Q132E, Q132F, Q132G, Q132H, Q132I, Q132K, Q132L, Q132M, Q132N, Q132P,Q132R, Q132S, Q132T, Q132V, Q132W, Q132Y, N133*, N133A, N133D, N133E,N133F, N133G, N133H, N133I, N133K, N133L, N133M, N133P, N133Q, N133R,N133S, N133T, N133V, N133W, N133Y, S134*, S134A, S134D, S134E, S134F,S134G, S134H, S134I, S134K, S134L, S134M, S134N, S134P, S134Q, S134R,S134T, S134V, S134W, S134Y, Q135*, Q135A, Q135D, Q135E, Q135F, Q135G,Q135H, Q135I, Q135K, Q135L, Q135M, Q135N, Q135P, Q135R, Q135S, Q135T,Q135V, Q135W, Q135Y, G136*, G136A, G136D, G136E, G136F, G136H, G136I,G136K, G136L, G136M, G136N, G136P, G136Q, G136R, G136S, G136T, G136V,G136W, G136Y, G137*, G137A, G137D, G137E, G137F, G137H, G137I, G137K,G137L, G137M, G137N, G137P, G137Q, G137R, G137S, G137T, G137V, G137W,G137Y, V138*, V138A, V138D, V138E, V138F, V138G, V138H, V138I, V138K,V138L, V138M, V138N, V138P, V138Q, V138R, V138S, V138T, V138W, V138Y,L139*, L139A, L139D, L139E, L139F, L139G, L139H, L139I, L139K, L139M,L139N, L139P, L139Q, L139R, L139S, L139T, L139V, L139W, L139Y, N140*,N140A, N140D, N140E, N140F, N140G, N140H, N140I, N140K, N140L, N140M,N140P, N140Q, N140R, N140S, N140T, N140V, N140W, N140Y, G141*, G141A,G141D, G141E, G141F, G141H, G141I, G141K, G141L, G141M, G141N, G141P,G141Q, G141R, G141S, G141T, G141V, G141W, G141Y, F142*, F142A, F142D,F142E, F142G, F142H, F142I, F142K, F142L, F142M, F142N, F142P, F142Q,F142R, F142S, F142T, F142V, F142W, F142Y, Y143*, Y143A, Y143D, Y143E,Y143F, Y143G, Y143H, Y143I, Y143K, Y143L, Y143M, Y143N, Y143P, Y143Q,Y143R, Y143S, Y143T, Y143V, Y143W, S144*, S144A, S144D, S144D, S144F,S144G, S144H, S144I, S144K, S144L, S144M, S144N, S144P, S144Q, S144R,S144T, S144V, S144W, S144Y, A145*, A145D, A145E, A145F, A145G, A145H,A145I, A145K, A145L, A145M, A145N, A145P, A145Q, A145R, A145S, A145T,A145V, A145W, A145Y, N146*, N146A, N146D, N146E, N146F, N146G, N146H,N146I, N146K, N146L, N146M, N146P, N146Q, N146R, N146S, N146T, N146V,N146W, N146Y, K147*, K147A, K147D, K147E, K147F, K147G, K147H, K147I,K147L, K147M, K147N, K147P, K147Q, K147R, K147S, K147T, K147V, K147W,K147Y, V148*, V148A, V148D, V148E, V148F, V148G, V148H, V148I, V148K,V148L, V148M, V148N, V148P, V148Q, V148R, V148S, V148T, V148W, V148Y,A149*, A149D, A149E, A149F, A149G, A149H, A149I, A149K, A149L, A149M,A149N, A149P, A149Q, A149R, A149S, A149T, A149V, A149W, A149Y, Q150*,Q150A, Q150D, Q150E, Q150F, Q150G, Q150H, Q150I, Q150K, Q150L, Q150M,Q150N, Q150P, Q150R, Q150S, Q150T, Q150V, Q150W, Q150Y, F151*, F151A,F151D, F151E, F151G, F151H, F151I, F151K, F151L, F151M, F151N, F151P,F151Q, F151R, F151S, F151T, F151V, F151W, F151Y, D152*, D152A, D152E,D152F, D152G, D152H, D152I, D152K, D152L, D152M, D152N, D152P, D152Q,D152R, D152S, D152T, D152V, D152W, D152Y, P153*, P153A, P153D, P153E,P153F, P153G, P153H, P153I, P153K, P153L, P153M, P153N, P153Q, P153R,P153S, P153T, P153V, P153W, P153Y, S154*, S154A, S154D, S154E, S154F,S154G, S154H, S154I, S154K, S154L, S154M, S154N, S154P, S154Q, S154R,S154T, S154V, S154W, S154Y, K155*, K155A, K155D, K155E, K155F, K155G,K155H, K155I, K155L, K155M, K155N, K155P, K155Q, K155R, K155S, K155T,K155V, K155W, K155Y, P156*, P156A, P156D, P156E, P156F, P156G, P156H,P156I, P156K, P156L, P156M, P156N, P156Q, P156R, P156S, P156T, P156V,P156W, P156Y, Q157*, Q157A, Q157D, Q157E, Q157F, Q157G, Q157H, Q157I,Q157K, Q157L, Q157M, Q157N, Q157P, Q157R, Q157S, Q157T, Q157V, Q157W,Q157Y, Q158*, Q158A, Q158D, Q158E, Q158F, Q158G, Q158H, Q158I, Q158K,Q158L, Q158M, Q158N, Q158P, Q158R, Q158S, Q158T, Q158V, Q158W, Q158Y,T159*, T159A, T159D, T159E, T159F, T159G, T159H, T159I, T159K, T159L,T159M, T159N, T159P, T159Q, T159R, T159S, T159V, T159W, T159Y, K160*,K160A, K160D, K160E, K160F, K160G, K160H, K160I, K160L, K160M, K160N,K160P, K160Q, K160R, K160S, K160T, K160V, K160W, K160Y, G161*, G161A,G161D, G161E, G161F, G161H, G161I, G161K, G161L, G161M, G161N, G161P,G161Q, G161R, G161S, G161T, G161V, G161W, G161Y, T162A, T162D, T162E,T162F, T162G, T162H, T162I, T162K, T162L, T162M, T162N, T162P, T162Q,T162R, T162S, T162T, T162V, T162W, T162Y, W163*, W163A, W163D, W163E,W163F, W163G, W163H, W163I, W163K, W163L, W163M, W163N, W163P, W163Q,W163R, W163S, W163T, W163V, W163Y, F164*, F164A, F164D, F164E, F164G,F164H, F164I, F164K, F164L, F164M, F164N, F164P, F164Q, F164R, F164S,F164T, F164V, F164W, F164Y, Q165*, Q165A, Q165D, Q165E, Q165F, Q165G,Q165H, Q165I, Q165K, Q165L, Q165M, Q165N, Q165P, Q165R, Q165S, Q165T,Q165V, Q165W, Q165Y, I166*, I166A, I166D, I166E, I166F, I166G, I166H,I166K, I166L, I166M, I166N, I166P, I166Q, I166R, I166S, I166T, I166V,I166W, I166Y, T167*, T167A, T167D, T167E, T167F, T167G, T167H, T167I,T167K, T167L, T167M, T167N, T167P, T167Q, T167R, T167S, T167V, T167W,T167Y, K168*, K168A, K168D, K168E, K168F, K168G, K168H, K168I, K168L,K168M, K168N, K168P, K168Q, K168R, K168S, K168T, K168V, K168W, K168Y,F169*, F169A, F169D, F169E, F169G, F169H, F169I, F169K, F169L, F169M,F169N, F169P, F169Q, F169R, F169S, F169T, F169V, F169W, F169Y, T170*,T170A, T170D, T170E, T170F, T170G, T170H, T170I, T170K, T170L, T170M,T170N, T170P, T170Q, T170R, T170S, T170V, T170W, T170Y, G171*, G171A,G171D, G171E, G171F, G171H, G171I, G171K, G171L, G171M, G171N, G171P,G171Q, G171R, G171S, G171T, G171V, G171W, G171Y, A172*, A172D, A172E,A172F, A172G, A172H, A172I, A172K, A172L, A172M, A172N, A172P, A172Q,A172R, A172S, A172T, A172V, A172W, A172Y, A173*, A173D, A173E, A173F,A173G, A173H, A173I, A173K, A173L, A173M, A173N, A173P, A173Q, A173R,A173S, A173T, A173V, A173W, A173Y, G174*, G174A, G174D, G174E, G174F,G174H, G174I, G174K, G174L, G174M, G174N, G174P, G174Q, G174R, G174S,G174T, G174V, G174W, G174Y, P175*, P175A, P175D, P175E, P175F, P175G,P175H, P175I, P175K, P175L, P175M, P175N, P175Q, P175R, P175S, P175T,P175V, P175W, P175Y, Y176*, Y176A, Y176D, Y176E, Y176F, Y176G, Y176H,Y176I, Y176K, Y176L, Y176M, Y176N, Y176P, Y176Q, Y176R, Y176S, Y176T,Y176V, Y176W, K178*, K178A, K178D, K178E, K178F, K178G, K178H, K178I,K178L, K178M, K178N, K178P, K178Q, K178R, K178S, K178T, K178V, K178W,K178Y, A179*, A179D, A179E, A179F, A179G, A179H, A179I, A179K, A179L,A179M, A179N, A179P, A179Q, A179R, A179S, A179T, A179V, A179W, A179Y,L180*, L180A, L180D, L180E, L180F, L180G, L180H, L180I, L180K, L180M,L180N, L180P, L180Q, L180R, L180S, L180T, L180V, L180W, L180Y, G181*,G181A, G181D, G181E, G181F, G181H, G181I, G181K, G181L, G181M, G181N,G181P, G181Q, G181R, G181S, G181T, G181V, G181W, G181Y, S182*, S182A,S182D, S182E, S182F, S182G, S182H, S182I, S182K, S182L, S182M, S182N,S182P, S182Q, S182R, S182T, S182V, S182W, S182Y, N183*, N183A, N183D,N183E, N183F, N183G, N183H, N183I, N183K, N183L, N183M, N183P, N183Q,N183R, N183S, N183T, N183V, N183W, N183Y, D184*, D184A, D184E, D184F,D184G, D184H, D184I, D184K, D184L, D184M, D184N, D184P, D184Q, D184R,D184S, D184T, D184V, D184W, D184Y, K185*, K185A, K185D, K185E, K185F,K185G, K185H, K185I, K185L, K185M, K185N, K185P, K185Q, K185R, K185S,K185T, K185V, K185W, K185Y, S186*, S186A, S186D, S186E, S186F, S186G,S186H, S186I, S186K, S186L, S186M, S186N, S186P, S186Q, S186R, S186T,S186V, S186W, S186Y, V187*, V187A, V187D, V187E, V187F, V187G, V187H,V187I, V187K, V187L, V187M, V187N, V187P, V187Q, V187R, V187S, V187T,V187W, V187Y, D189*, D189A, D189E, D189F, D189G, D189H, D189I, D189K,D189L, D189M, D189N, D189P, D189Q, D189R, D189S, D189T, D189V, D189W,D189Y, K190*, K190A, K190D, K190E, K190F, K190G, K190H, K190I, K190L,K190M, K190N, K190P, K190Q, K190R, K190S, K190T, K190V, K190W, K190Y,N191*, N191A, N191D, N191E, N191F, N191G, N191H, N191I, N191K, N191L,N191M, N191P, N191Q, N191R, N191S, N191T, N191V, N191W, N191Y, K192*,K192A, K192D, K192E, K192F, K192G, K192H, K192I, K192L, K192M, K192N,K192P, K192Q, K192R, K192S, K192T, K192V, K192W, K192Y, N193*, N193A,N193D, N193E, N193F, N193G, N193H, N193I, N193K, N193L, N193M, N193P,N193Q, N193R, N193S, N193T, N193V, N193W, N193Y, I194*, I194A, I194D,I194E, I194F, I194G, I194H, I194K, I194L, I194M, I194N, I194P, I194Q,I194R, I194S, I194T, I194V, I194W, I194Y, A195*, A195D, A195E, A195F,A195G, A195H, A195I, A195K, A195L, A195M, A195N, A195P, A195Q, A195R,A195S, A195T, A195V, A195W, A195Y, G196*, G196A, G196D, G196E, G196F,G196H, G196I, G196K, G196L, G196M, G196N, G196P, G196Q, G196R, G196S,G196T, G196V, G196W, G196Y, D197*, D197A, D197E, D197F, D197G, D197H,D197I, D197K, D197L, D197M, D197N, D197P, D197Q, D197R, D197S, D197T,D197V, D197W, D197Y, W198*, W198A, W198D, W198E, W198F, W198G, W198H,W198I, W198K, W198L, W198M, W198N, W198P, W198Q, W198R, W198S, W198T,W198V, W198Y, G199*, G199A, G199D, G199E, G199F, G199H, G199I, G199K,G199L, G199M, G199N, G199P, G199Q, G199R, G199S, G199T, G199V, G199W,G199Y, F200*, F200A, F200D, F200E, F200G, F200H, F200I, F200K, F200L,F200M, F200N, F200P, F200Q, F200R, F200S, F200T, F200V, F200W, F200Y,D201*, D201A, D201E, D201F, D201G, D201H, D201I, D201K, D201L, D201M,D201N, D201P, D201Q, D201R, D201S, D201T, D201V, D201W, D201Y, P202*,P202A, P202D, P202E, P202F, P202G, P202H, P202I, P202K, P202L, P202M,P202N, P202Q, P202R, P202S, P202T, P202V, P202W, P202Y, A203*, A203D,A203E, A203F, A203G, A203H, A203I, A203K, A203L, A203M, A203N, A203P,A203Q, A203R, A203S, A203T, A203V, A203W, A203Y, K204*, K204A, K204D,K204E, K204F, K204G, K204H, K204I, K204L, K204M, K204N, K204P, K204Q,K204R, K204S, K204T, K204V, K204W, K204Y, W205*, W205A, W205D, W205E,W205F, W205G, W205H, W205I, W205K, W205L, W205M, W205N, W205P, W205Q,W205R, W205S, W205T, W205V, W205Y, A206*, A206D, A206E, A206F, A206G,A206H, A206I, A206K, A206L, A206M, A206N, A206P, A206Q, A206R, A206S,A206T, A206V, A206W, A206Y, Y207*, Y207A, Y207D, Y207E, Y207F, Y207G,Y207H, Y207I, Y207K, Y207L, Y207M, Y207N, Y207P, Y207Q, Y207R, Y207S,Y207T, Y207V, Y207W, Q208*, Q208A, Q208D, Q208E, Q208F, Q208G, Q208H,Q208I, Q208K, Q208L, Q208M, Q208N, Q208P, Q208R, Q208S, Q208T, Q208V,Q208W, Q208Y, Y209*, Y209A, Y209D, Y209E, Y209F, Y209G, Y209H, Y209I,Y209K, Y209L, Y209M, Y209N, Y209P, Y209Q, Y209R, Y209S, Y209T, Y209V,Y209W, D210*, D210A, D210E, D210F, D210G, D210H, D210I, D210K, D210L,D210M, D210N, D210P, D210Q, D210R, D210S, D210T, D210V, D210W, D210Y,E211*, E211A, E211D, E211F, E211G, E211H, E211I, E211K, E211L, E211M,E211N, E211P, E211Q, E211R, E211S, E211T, E211V, E211W, E211Y, K212*,K212A, K212D, K212E, K212F, K212G, K212H, K212I, K212L, K212M, K212N,K212P, K212Q, K212R, K212S, K212T, K212V, K212W, K212Y, N213*, N213A,N213D, N213E, N213F, N213G, N213H, N213I, N213K, N213L, N213M, N213P,N213Q, N213R, N213S, N213T, N213V, N213W, N213Y, N214*, N214A, N214D,N214E, N214F, N214G, N214H, N214I, N214K, N214L, N214M, N214P, N214Q,N214R, N214S, N214T, N214V, N214W, N214Y, K215*, K215A, K215D, K215E,K215F, K215G, K215H, K215I, K215L, K215M, K215N, K215P, K215Q, K215R,K215S, K215T, K215V, K215W, K215Y, F216*, F216A, F216D, F216E, F216G,F216H, F216I, F216K, F216L, F216M, F216N, F216P, F216Q, F216R, F216S,F216T, F216V, F216W, F216Y, N217*, N217A, N217D, N217E, N217F, N217G,N217H, N217I, N217K, N217L, N217M, N217P, N217Q, N217R, N217S, N217T,N217V, N217W, N217Y, Y218*, Y218A, Y218D, Y218E, Y218F, Y218G, Y218H,Y218I, Y218K, Y218L, Y218M, Y218N, Y218P, Y218Q, Y218R, Y218S, Y218T,Y218V, Y218W, V219*, V219A, V219D, V219E, V219F, V219G, V219H, V219I,V219K, V219L, V219M, V219N, V219P, V219Q, V219R, V219S, V219T, V219W,V219Y, G220*, G220A, G220D, G220E, G220F, G220H, G220I, G220K, G220L,G220M, G220N, G220P, G220Q, G220R, G220S, G220T, G220V, G220W, G220Y,K221*, K221A, K221D, K221E, K221F, K221G, K221H, K221I, K221L, K221M,K221N, K221P, K221Q, K221R, K221S, K221T, K221V, K221W and K221Y and adetergent adjunct.

In one embodiment of the invention, the detergent adjunct ingredient isselected from the group consisting of surfactants, builders,flocculating aid, chelating agents, dye transfer inhibitors, enzymes,enzyme stabilizers, enzyme inhibitors, catalytic materials, bleachactivators, hydrogen peroxide, sources of hydrogen peroxide, preformedperacids, polymeric dispersing agents, clay soilremoval/anti-redeposition agents, brighteners, suds suppressors, dyes,perfumes, structure elasticizing agents, fabric softeners, carriers,hydrotropes, builders and co-builders, fabric hueing agents,anti-foaming agents, dispersants, processing aids, and/or pigments.

The detergent adjunct ingredient may be a surfactant. One advantage ofincluding a surfactant in a detergent composition comprising a DNasevariant is that the wash performance is improved. In one embodiment, thedetergent adjunct ingredient is a builder or a clay soilremoval/anti-redeposition agent.

In one embodiment, detergent adjunct ingredient is an enzyme. Thedetergent composition may comprise one or more enzymes, as specifiedbelow. The one or more enzymes may be selected from the group consistingof proteases, lipases, cutinases, amylases, carbohydrases, cellulases,pectinases, mannanases, arabinases, galactanases, xylanases andoxidases. Specific enzymes suitable for the detergent compositions ofthe invention are described below.

In one embodiment, the detergent composition comprising a DNase variantis capable of reducing adhesion of bacteria selected from the groupconsisting of Acinetobacter sp., Aeromicrobium sp., Brevundimonas sp.,Microbacterium sp., Micrococcus luteus, Pseudomonas sp., Staphylococcusepidermidis, and Stenotrophomonas sp. to a surface, or releasing thebacteria from a surface to which they adhere. Biofilm growth in laundryitems may originate from many organisms as described previously. Oneparticular abundant bacterium in biofilm originates from Brevundimonas.The DNase variants of the invention are particularly effective inreducing the growth of the bacterium and reducing the malodor,stickiness and re-deposition coursed by these bacteria. In oneembodiment of the invention, the surface is a textile surface. Thetextile can be made of cotton, Cotton/Polyester, Polyester, Polyamide,Polyacryl and/or silk.

The detergent composition may be formulated as a bar, a homogenoustablet, and a tablet having two or more layers, a pouch having one ormore compartments, a regular or compact powder, a granule, a paste, agel, or a regular, compact or concentrated liquid. The detergentcomposition can be a liquid detergent, a powder detergent or a granuledetergent.

The DNase variants of the invention are suitable for use in cleaningsuch as laundry. The invention further relates a method for launderingan item, which method comprises the steps of:

a. Exposing an item to a wash liquor comprising a DNase variant of theinvention;

b. Completing at least one wash cycle; and

c. Optionally rinsing the item,

wherein the item is a textile.

The pH of the liquid solution is in the range of 1 to 11, such as in therange 5.5 to 11, such as in the range of 7 to 9, in the range of 7 to 8or in the range of 7 to 8.5.

The wash liquor may have a temperature in the range of 5° C. to 95° C.,or in the range of 10° C. to 80° C., in the range of 10° C. to 70° C.,in the range of 10° C. to 60° C., in the range of 10° C. to 50° C., inthe range of 15° C. to 40° C. or in the range of 20° C. to 30° C. In oneembodiment the temperature of the wash liquor is 30° C.

In one embodiment of the invention, the method for laundering an itemfurther comprises draining of the wash liquor or part of the wash liquorafter completion of a wash cycle. The wash liquor can then be re-used ina subsequent wash cycle or in a subsequent rinse cycle. The item may beexposed to the wash liquor during a first and optionally a second or athird wash cycle. In one embodiment the item is rinsed after beingexposed to the wash liquor. The item can be rinsed with water or withwater comprising a conditioner. The invention further concerns an itemwashed according to the method.

The DNases of the invention may be added to a wash liquor.

The concentration of the DNase variants in the wash liquor is typicallyin the range of 0.00004-100 ppm enzyme protein, such as in the range of0.00008-100, in the range of 0.0001-100, in the range of 0.0002-100, inthe range of 0.0004-100, in the range of 0.0008-100, in the range of0.001-100 ppm enzyme protein, 0.01-100 ppm enzyme protein, preferably0.05-50 ppm enzyme protein, more preferably 0.1-50 ppm enzyme protein,more preferably 0.1-30 ppm enzyme protein, more preferably 0.5-20 ppmenzyme protein, and most preferably 0.5-10 ppm enzyme protein.

The DNase variant of the present invention may be added to a detergentcomposition in an amount corresponding to at least 0.002 mg of DNaseprotein, such as at least 0.004 mg of DNase protein, at least 0.006 mgof DNase protein, at least 0.008 mg of DNase protein, at least 0.01 mgof DNase protein, at least 0.1 mg of protein, preferably at least 1 mgof protein, more preferably at least 10 mg of protein, even morepreferably at least 15 mg of protein, most preferably at least 20 mg ofprotein, and even most preferably at least 25 mg of protein. Thus, thedetergent composition may comprise at least 0.00008% DNase protein,preferably at least 0.002%, 0.003%, 0.004%, 0.005%, 0.006%, 0.008%,0.01%, 0.02%, 0.03%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, 0.6%, 0.7%, 0.8%,0.9% or 1.0% of DNase protein.

Liquid Detergent Composition

The DNase variants of the invention may also be formulated in liquidlaundry compositions such as a liquid laundry compositions compositioncomprising:

a) at least 0.005 mg of active DNase variant protein per litredetergent,

b) 2 wt % to 60 wt % of at least one surfactant, and

c) 5 wt % to 50 wt % of at least one builder

The surfactant may be selected among nonionic, anionic and/or amphotericsurfactants as described above, preferably anionic or nonionicsurfactants but also amphoteric surfactants may be used. In general,bleach-stable surfactants are preferred. Preferred anionic surfactantsare sulphate surfactants and in particular alkyl ether sulphates,especially C-9-15 alcohol ethersulfates, C12-15 primary alcoholethoxylate, C8-C16 ester sulphates and C10-C14 ester sulphates, such asmono dodecyl ester sulphates Non-limiting examples of anionicsurfactants include sulfates and sulfonates, in particular, linearalkylbenzenesulfonates (LAS), isomers of LAS, branchedalkylbenzenesulfonates (BABS), phenylalkanesulfonates,alpha-olefinsulfonates (AOS), olefin sulfonates, alkene sulfonates,alkane-2,3-diylbis(sulfates), hydroxyalkanesulfonates and disulfonates,alkyl sulfates (AS) such as sodium dodecyl sulfate (SDS), fatty alcoholsulfates (FAS), primary alcohol sulfates (PAS), alcohol ethersulfates(AES or AEOS or FES, also known as alcohol ethoxysulfates or fattyalcohol ether sulfates), secondary alkanesulfonates (SAS), paraffinsulfonates (PS), ester sulfonates, sulfonated fatty acid glycerolesters, alpha-sulfo fatty acid methyl esters (alpha-SFMe or SES)including methyl ester sulfonate (MES), alkyl- or alkenylsuccinic acid,dodecenyl/tetradecenyl succinic acid (DTSA), fatty acid derivatives ofamino acids, diesters and monoesters of sulfo-succinic acid or salt offatty acids (soap), and combinations thereof.

The anionic surfactants are preferably added to the detergent in theform of salts. Suitable cations in these salts are alkali metal ions,such as sodium, potassium and lithium and ammonium salts, for example(2-hydroxyethyl)ammonium, bis(2-hydroxyethyl)ammonium andtris(2-hydroxyethyl)ammonium salts. Non-limiting examples of nonionicsurfactants include alcohol ethoxylates (AE or AEO), alcoholpropoxylates, propoxylated fatty alcohols (PFA), alkoxylated fatty acidalkyl esters, such as ethoxylated and/or propoxylated fatty acid alkylesters, alkylphenol ethoxylates (APE), nonylphenol ethoxylates (NPE),alkylpolyglycosides (APG), alkoxylated amines, fatty acidmonoethanolamides (FAM), fatty acid diethanolamides (FADA), ethoxylatedfatty acid monoethanolamides (EFAM), propoxylated fatty acidmonoethanolamides (PFAM), polyhydroxyalkyl fatty acid amides, or N-acylN-alkyl derivatives of glucosamine (glucamides, GA, or fatty acidglucamides, FAGA), as well as products available under the trade namesSPAN and TWEEN, and combinations thereof. Commercially availablenonionic surfactants includes Plurafac™, Lutensol™ and Pluronic™ fromBASF, Dehypon™ series from Cognis and Genapol™ series from Clariant.

The builder is preferably selected among phosphates, sodium citratebuilders, sodium carbonate, sodium silicate, sodium aluminosilicate(zeolite). Suitable builders are alkali metal or ammonium phosphates,polyphosphates, phosphonates, polyphosphates, carbonates, bicarbonates,borates, citrates, and polycarboxylates. Citrate builders, e.g., citricacid and soluble salts thereof (particularly sodium salt), arepolycarboxylate builders. Citrates can be used in combination withzeolite, silicates like the BRITESIL types, and/or layered silicatebuilders. The builder is preferably added in an amount of about 0-65% byweight, such as about 5% to about 50% by weight. In a laundry detergent,the level of builder is typically about 40-65% by weight, particularlyabout 50-65% by weight, particularly from 20% to 50% by weight. Thebuilder and/or co-builder may particularly be a chelating agent thatforms water-soluble complexes with Ca and Mg. Any builder and/orco-builder known in the art for use in cleaning detergents may beutilized. Non-limiting examples of builders include zeolites,diphosphates (pyrophosphates), triphosphates such as sodium triphosphate(STP or STPP), carbonates such as sodium carbonate, soluble silicatessuch as sodium metasilicate, layered silicates (e.g., SKS-6 fromHoechst), and (carboxymethyl)inulin (CMI), and combinations thereof.Further non-limiting examples of builders include citrate, chelatorssuch as aminocarboxylates, aminopolycarboxylates and phosphonates, andalkyl- or alkenylsuccinic acid. Additional specific examples include2,2′,2″-nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid(EDTA), diethylenetriaminepentaacetic acid (DTPA), iminodisuccinic acid(IDS), ethylenediamine-N,N′-disuccinic acid (EDDS),methylglycine-N,N-diacetic acid (MGDA), glutamic acid-N,N-diacetic acid(GLDA), 1-hydroxyethane-1,1-diphosphonic acid,N-(2-hydroxyethyl)iminodiacetic acid (EDG), aspartic acid-N-monoaceticacid (ASMA), aspartic acid-N,N-diacetic acid (ASDA), asparticacid-N-monopropionic acid (ASMP), iminodisuccinic acid (IDA),N-(sulfomethyl)aspartic acid (SMAS), N-(2-sulfoethyl)-aspartic acid(SEAS), N-(sulfomethylglutamic acid (SMGL), N-(2-sulfoethyl)-glutamicacid (SEGL), N-methyliminodiacetic acid (MIDA), serine-N,N-diacetic acid(SEDA), isoserine-N,N-diacetic acid (ISDA), phenylalanine-N,N-diaceticacid (PHDA), anthranilic acid-N,N-diacetic acid (ANDA), sulfanilicacid-N,N-diacetic acid (SLDA), taurine-N,N-diacetic acid (TUDA) andN′-(2-hydroxyethyl)ethylenediamine-N,N,N′-triacetic acid (HEDTA),diethanolglycine (DEG), and combinations and salts thereof. Phosphonatessuitable for use herein include 1-hydroxyethane-1,1-diphosphonic acid(HEDP), ethylenediaminetetrakis (methylenephosphonicacid) (EDTMPA),diethylenetriaminepentakis(methylenephosphonic acid) (DTMPA or DTPMPA orDTPMP), nitrilotris(methylenephosphonic acid) (ATMP or NTMP),2-phosphonobutane-1,2,4-tricarboxylic acid (PBTC),hexamethylenediaminetetrakis(methylenephosphonic acid) (HDTMP).

The composition may also contain 0-50% by weight, such as about 5% toabout 30%, of a detergent co-builder. The detergent composition mayinclude a co-builder alone, or in combination with a builder, forexample a zeolite builder. Non-limiting examples of co-builders includehomopolymers of polyacrylates or copolymers thereof, such aspoly(acrylic acid) (PAA) or copoly(acrylic acid/maleic acid) (PAA/PMA)or polyaspartic acid. Further exemplary builders and/or co-builders aredescribed in, e.g., WO 09/102854, U.S. Pat. No. 5,977,053

In one preferred embodiment, the builder is a non-phosphorus basedbuilder such as citric acid and/or methylglycine-N,N-diacetic acid(MGDA) and/or glutamic-N,N-diacetic acid (GLDA) and/or salts thereof.

The laundry composition may also be phosphate free in the instance thepreferred builders includes citrate and/or methylglycine-N,N-diaceticacid (MGDA) and/or glutamic-N,N-diacetic acid (GLDA) and/or saltsthereof.

One embodiment of the invention concerns a liquid laundry compositionscomposition comprising:

a) at least 0.005 mg of DNase variant per litre of composition,

b) 1% to 15% by weight of at least one surfactant wherein the surfactantis LAS, AEOS and/or SLES, and

c) 5% to 50% by weight of at least one builder selected from HEDP, DTMPAor DTPMPA.

The liquid detergent composition may typically containing at least 20%by weight and up to 95% water, such as up to 70% water, up to 50% water,up to 40% water, up to 30% water, or up to 20% water. Other types ofliquids, including without limitation, alkanols, amines, diols, ethersand polyols may be included in an aqueous liquid detergent. An aqueousliquid detergent may contain from 0-30% organic solvent. A liquiddetergent may even be non-aqueous, wherein the water content is below10%, preferably below 5%.

Powder Compositions

The detergent composition may also be formulated into a granulardetergent for laundry or dish wash. One embodiment of the inventionconcerns a granular detergent composition comprising

a) at least 0.005 mg of DNase variant per gram of composition,

b) 5 wt % to 50 wt % anionic surfactant

c) 1 wt % to 8 wt % nonionic surfactant, and

d) 5 wt % to 40 wt % builder such as carbonates, zeolites, phosphatebuilder, calcium sequestering builders or complexing agents

The surfactant may be selected among nonionic, anionic and/or amphotericsurfactants as described above, preferably anionic or nonionicsurfactants but also amphoteric surfactants may be used. In general,bleach-stable surfactants are preferred. Preferred anionic surfactantsare sulphate surfactants and in particular alkyl ether sulphates,especially C-9-15 alcohol ethersulfates, C12-15 primary alcoholethoxylate, C8-C16 ester sulphates and C10-C14 ester sulphates, such asmono dodecyl ester sulphates Non-limiting examples of anionicsurfactants include sulfates and sulfonates, in particular, linearalkylbenzenesulfonates (LAS), isomers of LAS, branchedalkylbenzenesulfonates (BABS), phenylalkanesulfonates,alpha-olefinsulfonates (AOS), olefin sulfonates, alkene sulfonates,alkane-2,3-diylbis(sulfates), hydroxyalkanesulfonates and disulfonates,alkyl sulfates (AS) such as sodium dodecyl sulfate (SDS), fatty alcoholsulfates (FAS), primary alcohol sulfates (PAS), alcohol ethersulfates(AES or AEOS or FES, also known as alcohol ethoxysulfates or fattyalcohol ether sulfates), secondary alkanesulfonates (SAS), paraffinsulfonates (PS), ester sulfonates, sulfonated fatty acid glycerolesters, alpha-sulfo fatty acid methyl esters (alpha-SFMe or SES)including methyl ester sulfonate (MES), alkyl- or alkenylsuccinic acid,dodecenyl/tetradecenyl succinic acid (DTSA), fatty acid derivatives ofamino acids, diesters and monoesters of sulfo-succinic acid or salt offatty acids (soap), and combinations thereof. The anionic surfactantsare preferably added to the detergent in the form of salts. Suitablecations in these salts are alkali metal ions, such as sodium, potassiumand lithium and ammonium salts, for example (2-hydroxyethyl)ammonium,bis(2-hydroxyethyl)ammonium and tris(2-hydroxyethyl)ammonium salts.

Non-limiting examples of nonionic surfactants include alcoholethoxylates (AE or AEO), alcohol propoxylates, propoxylated fattyalcohols (PFA), alkoxylated fatty acid alkyl esters, such as ethoxylatedand/or propoxylated fatty acid alkyl esters, alkylphenol ethoxylates(APE), nonylphenol ethoxylates (NPE), alkylpolyglycosides (APG),alkoxylated amines, fatty acid monoethanolamides (FAM), fatty aciddiethanolamides (FADA), ethoxylated fatty acid monoethanolamides (EFAM),propoxylated fatty acid monoethanolamides (PFAM), polyhydroxyalkyl fattyacid amides, or N-acyl N-alkyl derivatives of glucosamine (glucamides,GA, or fatty acid glucamides, FAGA), as well as products available underthe trade names SPAN and TWEEN, and combinations thereof.

Commercially available nonionic surfactants includes Plurafac™,Lutensol™ and Pluronic™ range from BASF, Dehypon™ series from Cognis andGenapol™ series from Clariant.

The builder is may be non-phosphate such as citrate preferably as asodium salt and/or zeolites. Phosphonate builder may be any of thosedescribed above.

The builder is preferably selected among phosphates and sodium citratebuilders, sodium carbonate, sodium silicate, sodium aluminosilicate(zeolite) as described above. Suitable builders are described above andinclude alkali metal or ammonium phosphates, polyphosphates,phosphonates, polyphosphonates, carbonates, bicarbonates, borates,polyhydroxysulfonates, polyacetates, carboxylates, citrates, andpolycarboxylates. Citrate builders, e.g., citric acid and soluble saltsthereof (particularly sodium salt), are polycarboxylate builders. Thebuilder is preferably added in an amount of about 0-65% by weight, suchas about 5% to about 50% by weight, such as 5 to 40% by weight, such as10 to 40% by weight, such as 10 to 30% by weight, such as 15 to 20% byweight or such as 20 to 40% by weight. The builder may be a phosphonatebuilder including 1-hydroxyethane-1,1-diphosphonic acid (HEDP),ethylenediaminetetra (methylenephosphonic acid) (EDTMPA),diethylenetriaminepentakis (methylenephosphonic acid) (DTMPA or DTPMPA),diethylenetriamine penta (methylenephosphonic acid) (DTPMP),aminotris(methylenephosphonic acid) (ATMP),2-phosphonobutane-1,2,4-tricarboxylic acid (PBTC) andhexamethylenediaminetetra (methylenephosphonic acid) (HDTMP). Preferredphosphonates includes 1-hydroxyethane-1,1-diphosphonic acid (HEDP)and/or diethylenetriaminepentakis (methylenephosphonic acid) (DTMPA orDTPMPA). The phosphonate is preferably added in an amount of about in alevel of from about 0.01% to about 10% by weight, preferably from 0.1%to about 5% by weight, more preferably from 0.5% to 3% by weight of thecomposition.

The laundry composition may also be phosphate free in the instance thepreferred builders includes citrate, carbonates and/or sodiumaluminosilicate (zeolite).

The detergent may contain 0-30% by weight, such as about 1% to about20%, of a bleaching system. Any bleaching system comprising componentsknown in the art for use in cleaning detergents may be utilized.Suitable bleaching system components include sources of hydrogenperoxide; sources of peracids; and bleach catalysts or boosters.

Sources of hydrogen peroxide: Suitable sources of hydrogen peroxide areinorganic persalts, including alkali metal salts such as sodiumpercarbonate and sodium perborates (usually mono- or tetrahydrate), andhydrogen peroxide-urea (1/1).

Sources of peracids: Peracids may be (a) incorporated directly aspreformed peracids or (b) formed in situ in the wash liquor fromhydrogen peroxide and a bleach activator (perhydrolysis) or (c) formedin situ in the wash liquor from hydrogen peroxide and a perhydrolase anda suitable substrate for the latter, e.g., an ester.

a) Suitable preformed peracids include, but are not limited to,peroxycarboxylic acids such as peroxybenzoic acid and itsring-substituted derivatives, peroxy-α-naphthoic acid, peroxyphthalicacid, peroxylauric acid, peroxystearic acid, ε-phthalimidoperoxycaproicacid [phthalimidoperoxyhexanoic acid (PAP)], ando-carboxybenzamidoperoxycaproic acid; aliphatic and aromaticdiperoxydicarboxylic acids such as diperoxydodecanedioic acid,diperoxyazelaic acid, diperoxysebacic acid, diperoxybrassylic acid,2-decyldiperoxybutanedioic acid, and diperoxyphthalic, -isophthalic and-terephthalic acids; perimidic acids; peroxymonosulfuric acid;peroxydisulfuric acid; peroxyphosphoric acid; peroxysilicic acid; andmixtures of said compounds. It is understood that the peracids mentionedmay in some cases be best added as suitable salts, such as alkali metalsalts (e.g., Oxone®) or alkaline earth-metal salts.

b) Suitable bleach activators include those belonging to the class ofesters, amides, imides, nitriles or anhydrides and, where applicable,salts thereof. Suitable examples are tetraacetylethylenediamine (TAED),sodium 4-[(3,5,5-trimethylhexanoyl)oxy]benzene-1-sulfonate (ISONOBS),sodium 4-(dodecanoyloxy)benzene-1-sulfonate (LOBS), sodium4-(decanoyloxy)benzene-1-sulfonate, 4-(decanoyloxy)benzoic acid (DOBA),sodium 4-(nonanoyloxy)benzene-1-sulfonate (NOBS), and/or those disclosedin WO98/17767. A particular family of bleach activators of interest wasdisclosed in EP624154 and particularly preferred in that family isacetyl triethyl citrate (ATC). ATC or a short chain triglyceride liketriacetin has the advantage that they are environmentally friendly.Furthermore acetyl triethyl citrate and triacetin have good hydrolyticalstability in the product upon storage and are efficient bleachactivators. Finally ATC is multifunctional, as the citrate released inthe perhydrolysis reaction may function as a builder.

Bleach catalysts and boosters: The bleaching system may also include ableach catalyst or booster. Some non-limiting examples of bleachcatalysts that may be used in the compositions of the present inventioninclude manganese oxalate, manganese acetate, manganese-collagen,cobalt-amine catalysts and manganese triazacyclononane (MnTACN)catalysts; particularly preferred are complexes of manganese with1,4,7-trimethyl-1,4,7-triazacyclononane (Me3-TACN) or1,2,4,7-tetramethyl-1,4,7-triazacyclononane (Me4-TACN), in particularMe3-TACN, such as the dinuclear manganese complex[(Me3-TACN)Mn(O)3Mn(Me3-TACN)](PF6)2, and[2,2′,2″-nitrilotris(ethane-1,2-diylazanylylidene-κN-methanylylidene)triphenolato-κ30]manganese(III).The bleach catalysts may also be other metal compounds, such as iron orcobalt complexes.

In some embodiments, where a source of a peracid is included, an organicbleach catalyst or bleach booster may be used having one of thefollowing formulae:

(iii) and mixtures thereof; wherein each R1 is independently a branchedalkyl group containing from 9 to 24 carbons or linear alkyl groupcontaining from 11 to 24 carbons, preferably each R1 is independently abranched alkyl group containing from 9 to 18 carbons or linear alkylgroup containing from 11 to 18 carbons, more preferably each R1 isindependently selected from the group consisting of 2-propylheptyl,2-butyloctyl, 2-pentylnonyl, 2-hexyldecyl, dodecyl, tetradecyl,hexadecyl, octadecyl, isononyl, isodecyl, isotridecyl and isopentadecyl.

Other exemplary bleaching systems are described, e.g. in WO 2007/087258,WO 2007/087244, WO 2007/087259, EP 1867708 (Vitamin K) and WO2007/087242. Suitable photobleaches may for example be sulfonated zincor aluminium phthalocyanines.

According to one embodiment and any of the previous embodiments theinvention also relates to a detergent composition comprising;

a) at least 0.005 mg of DNase variant per gram of composition,

b) 10-50 wt % builder and

c) at least one bleach component, wherein the bleach is a peroxide andthe bleach catalyst is a manganese compound.

The oxygen bleach is preferably percarbonate and the manganese catalystpreferably 1,4,7-trimethyl-1,4,7-triazacyclononane or manganese (III)acetate tetrahydrate

According to one embodiment and any of the previous embodiments theinvention also relates to a cleaning composition comprising;

a) at least 0.005 mg of active DNase variant,

b) 10-50 wt % builder selected from citric acid, methylglycine-N,N-diacetic acid (MGDA) and/or glutamic-N,N-diacetic acid(GLDA) and mixtures thereof, and

c) at least one bleach component, wherein the bleach is an oxygen bleachand the bleach catalyst is a manganese compound.

The oxygen bleach is preferably percarbonate and the manganese catalystpreferably 1,4,7-trimethyl-1,4,7-triazacyclo-nonane or manganese (II)acetate tetrahydrate

According to one embodiment and any of the previous embodiments theinvention also relates to a detergent composition comprising;

a) at least 0.005 mg of active DNase variant per gram of composition,

b) 10-50 wt % builder selected from citric acid, methylglycine-N,N-diacetic acid (MGDA) and/or glutamic-N,N-diacetic acid(GLDA) and mixtures thereof, and

c) 0.1-40 wt %, preferably from 0.5-30 wt %, of bleaching components,wherein the bleach components are a peroxide, preferably percabonate anda metal-containing bleach catalyst preferably1,4,7-trimethyl-1,4,7-triazacyclononane or manganese (II) acetatetetrahydrate (MnTACN).

The choice of detergent components may include, for textile care, theconsideration of the type of textile to be cleaned, the type and/ordegree of soiling, the temperature at which cleaning is to take place,and the formulation of the detergent product. Although componentsmentioned below are categorized by general header according to aparticular functionality, this is not to be construed as a limitation,as a component may comprise additional functionalities as will beappreciated by the skilled artisan, including the exemplary non-limitingcomponents set forth below.

Hydrotropes

The detergent composition may contain 0-10% by weight, for example 0-5%by weight, such as about 0.5 to about 5%, or about 3% to about 5%, of ahydrotrope. Any hydrotrope known in the art for use in detergents may beutilized. Non-limiting examples of hydrotropes include sodiumbenzenesulfonate, sodium p-toluene sulfonate (STS), sodium xylenesulfonate (SXS), sodium cumene sulfonate (SCS), sodium cymene sulfonate,amine oxides, alcohols and polyglycolethers, sodium hydroxynaphthoate,sodium hydroxynaphthalene sulfonate, sodium ethylhexyl sulfate, andcombinations thereof.

Polymers

The detergent composition may contain 0-10% by weight, such as 0.5-5%,2-5%, 0.5-2% or 0.2-1% of a polymer. Any polymer known in the art foruse in detergents may be utilized. The polymer may function as aco-builder as mentioned above, or may provide antiredeposition, fibreprotection, soil release, dye transfer inhibition, grease cleaningand/or anti-foaming properties. Some polymers may have more than one ofthe above-mentioned properties and/or more than one of thebelow-mentioned motifs. Exemplary polymers include(carboxymethyl)cellulose (CMC), poly(vinyl alcohol) (PVA),poly(vinylpyrrolidone) (PVP), poly(ethyleneglycol) or poly(ethyleneoxide) (PEG), ethoxylated poly(ethyleneimine), carboxymethyl inulin(CMI), and polycarboxylates such as PAA, PAA/PMA, polyaspartic acid, andlauryl methacrylate/acrylic acid copolymers, hydrophobically modifiedCMC (HM-CMC) and silicones, copolymers of terephthalic acid andoligomeric glycols, copolymers of poly(ethylene terephthalate) andpoly(oxyethene terephthalate) (PET-POET), PVP, poly(vinylimidazole)(PVI), poly(vinylpyridine-N-oxide) (PVPO or PVPNO) andpolyvinylpyrrolidone-vinylimidazole (PVPVI). Further exemplary polymersinclude sulfonated polycarboxylates, polyethylene oxide andpolypropylene oxide (PEO-PPO) and diquaternium ethoxy sulfate. Otherexemplary polymers are disclosed in, e.g., WO 2006/130575. Salts of theabove-mentioned polymers are also contemplated.

Fabric Hueing Agents

The detergent composition of the present invention may also includefabric hueing agents such as dyes or pigments, which when formulated indetergent compositions can deposit onto a fabric when said fabric iscontacted with a wash liquor comprising said detergent compositions andthus altering the tint of said fabric through absorption/reflection ofvisible light. Fluorescent whitening agents emit at least some visiblelight. In contrast, fabric hueing agents alter the tint of a surface asthey absorb at least a portion of the visible light spectrum. Suitablefabric hueing agents include dyes and dye-clay conjugates, and may alsoinclude pigments. Suitable dyes include small molecule dyes andpolymeric dyes. Suitable small molecule dyes include small molecule dyesselected from the group consisting of dyes falling into the Colour Index(C.I.) classifications of Direct Blue, Direct Red, Direct Violet, AcidBlue, Acid Red, Acid Violet, Basic Blue, Basic Violet and Basic Red, ormixtures thereof, for example as described in WO 2005/03274, WO2005/03275, WO 2005/03276 and EP 1876226 (hereby incorporated byreference). The detergent composition preferably comprises from about0.00003 wt % to about 0.2 wt %, from about 0.00008 wt % to about 0.05 wt%, or even from about 0.0001 wt % to about 0.04 wt % fabric hueingagent. The composition may comprise from 0.0001 wt % to 0.2 wt % fabrichueing agent, this may be especially preferred when the composition isin the form of a unit dose pouch. Suitable hueing agents are alsodisclosed in, e.g. WO 2007/087257 and WO 2007/087243.

Enzymes

The detergent composition may comprise one or more additional enzymessuch as a protease, lipase, cutinase, an amylase, carbohydrase,cellulase, pectinase, mannanase, arabinase, galactanase, xylanase,oxidase, e.g., a laccase, and/or peroxidase.

In general, the properties of the selected enzyme(s) should becompatible with the selected detergent, (i.e., pH-optimum, compatibilitywith other enzymatic and non-enzymatic ingredients, etc.), and theenzyme(s) should be present in effective amounts.

Cellulases

Suitable cellulases include those of bacterial or fungal origin.Chemically modified or protein engineered mutants are included. Suitablecellulases include cellulases from the genera Bacillus, Pseudomonas,Humicola, Fusarium, Thielavia, Acremonium, e.g., the fungal cellulasesproduced from Humicola insolens, Myceliophthora thermophila and Fusariumoxysporum disclosed in U.S. Pat. Nos. 4,435,307, 5,648,263, 5,691,178,5,776,757 and WO 89/09259.

Especially suitable cellulases are the alkaline or neutral cellulaseshaving colour care benefits. Examples of such cellulases are cellulasesdescribed in EP 0 495 257, EP 0 531 372, WO 96/11262, WO 96/29397, WO98/08940. Other examples are cellulase variants such as those describedin WO 94/07998, EP 0 531 315, U.S. Pat. Nos. 5,457,046, 5,686,593,5,763,254, WO 95/24471, WO 98/12307 and WO 99/001544.

Other cellulases are endo-beta-1,4-glucanase enzyme having a sequence ofat least 97% identity to the amino acid sequence of position 1 toposition 773 of SEQ ID NO: 2 of WO 2002/099091 or a family 44xyloglucanase, which a xyloglucanase enzyme having a sequence of atleast 60% identity to positions 40-559 of SEQ ID NO: 2 of WO2001/062903.

Commercially available cellulases include Celluzyme™, and Carezyme™(Novozymes A/S) Carezyme Premium™ (Novozymes A/S), Celluclean™(Novozymes A/S), Celluclean Classic™ (Novozymes A/S), Cellusoft™(Novozymes A/S), Whitezyme™ (Novozymes A/S), Clazinase™, and Puradax HA™(Genencor International Inc.), and KAC-500(B)™ (Kao Corporation).

Proteases

Suitable proteases include those of bacterial, fungal, plant, viral oranimal origin e.g. vegetable or microbial origin. Microbial origin ispreferred. Chemically modified or protein engineered mutants areincluded. It may be an alkaline protease, such as a serine protease or ametalloprotease. A serine protease may for example be of the S1 family,such as trypsin, or the S8 family such as subtilisin. A metalloproteasesprotease may for example be a thermolysin from e.g. family M4 or othermetalloprotease such as those from M5, M7 or M8 families.

The term “subtilases” refers to a sub-group of serine protease accordingto Siezen et al., Protein Engng. 4 (1991) 719-737 and Siezen et al.Protein Science 6 (1997) 501-523. Serine proteases are a subgroup ofproteases characterized by having a serine in the active site, whichforms a covalent adduct with the substrate. The subtilases may bedivided into 6 sub-divisions, i.e. the Subtilisin family, the Thermitasefamily, the Proteinase K family, the Lantibiotic peptidase family, theKexin family and the Pyrolysin family.

Examples of subtilases are those obtained from Bacillus such as Bacilluslentus, B. alkalophilus, B. subtilis, B. amyloliquefaciens, Bacilluspumilus and Bacillus gibsonii described in; U.S. Pat. No. 7,262,042 andWO 09/021867, and subtilisin lentus, subtilisin Novo, subtilisinCarlsberg, Bacillus licheniformis, subtilisin BPN′, subtilisin 309,subtilisin 147 and subtilisin 168 described in WO 89/06279 and proteasePD138 described in (WO 93/18140). Other useful proteases may be thosedescribed in WO 92/175177, WO 01/016285, WO 02/026024 and WO 02/016547.Examples of trypsin-like proteases are trypsin (e.g. of porcine orbovine origin) and the Fusarium protease described in WO 89/06270, WO94/25583 and WO 05/040372, and the chymotrypsin proteases obtained fromCellulomonas described in WO 05/052161 and WO 05/052146.

A further preferred protease is the alkaline protease from Bacilluslentus DSM 5483, as described for example in WO 95/23221, and variantsthereof which are e.g. described in WO 92/21760, WO 95/23221, EP 1921147and EP 1921148.

Examples of metalloproteases are the neutral metalloprotease asdescribed in WO 07/044993 (Genencor Int.) such as those obtained fromBacillus amyloliquefaciens.

Examples of useful proteases are the variants described in: WO 92/19729,WO 96/034946, WO 98/20115, WO 98/20116, WO 99/011768, WO 01/44452, WO03/006602, WO 04/03186, WO 04/041979, WO 07/006305, WO 11/036263, WO11/036264, especially the variants with substitutions in one or more ofthe following positions: 3, 4, 9, 15, 27, 36, 57, 68, 76, 87, 95, 96,97, 98, 99, 100, 101, 102, 103, 104, 106, 118, 120, 123, 128, 129, 130,160, 167, 170, 194, 195, 199, 205, 206, 217, 218, 222, 224, 232, 235,236, 245, 248, 252 and 274 using the BPN′ numbering. More preferred thesubtilase variants may comprise the mutations: S3T, V4I, S9R, A15T,K27R, *36D, V68A, N76D, N87S,R, *97E, A98S, S99G,D,A, S99AD, S101G,M,RS103A, V104I,Y,N, S106A, G118V,R, H120D,N, N123S, S128L, P129Q, S130A,G160D, Y167A, R170S, A194P, G195E, V199M, V205I, L217D, N218D, M222S,A232V, K235L, Q236H, Q245R, N252K, T274A (using BPN′ numbering).

Suitable commercially available protease enzymes include those soldunder the trade names Alcalase®, Duralase™, Durazym™, Relase®, Relase®Ultra, Savinase®, Savinase® Ultra, Primase®, Polarzyme®, Kannase®,Liquanase®, Liquanase® Ultra, Ovozyme®, Coronase®, Coronase® Ultra,Neutrase®, Everlase® and Esperase® (Novozymes A/S), those sold under thetradename Maxatase®, Maxacal®, Maxapem®, Purafect®, Purafect Prime®,Preferenz™, Purafect MA®, Purafect Ox®, Purafect OxP®, Puramax®,Properase®, Effectenz™, FN2®, FN3®, FN4®, Excellase®, Opticlean® andOptimase® (Danisco/DuPont), Axapem™ (Gist-Brocases N.V.), BLAP (sequenceshown in FIG. 29 of U.S. Pat. No. 5,352,604) and variants hereof (HenkelAG) and KAP (Bacillus alkalophilus subtilisin) from Kao.

Lipases and Cutinases

Suitable lipases and cutinases include those of bacterial or fungalorigin. Chemically modified or protein engineered mutant enzymes areincluded. Examples include lipase from Thermomyces, e.g. from T.lanuginosus (previously named Humicola lanuginosa) as described in EP258068 and EP 305216, cutinase from Humicola, e.g. H. insolens (WO96/13580), lipase from strains of Pseudomonas (some of these now renamedto Burkholderia), e.g. P. alcaligenes or P. pseudoalcaligenes (EP218272), P. cepacia (EP 331376), P. sp. strain SD705 (WO 95/06720 & WO96/27002), P. wisconsinensis (WO 96/12012), GDSL-type Streptomyceslipases (WO 10/065455), cutinase from Magnaporthe grisea (WO 10/107560),cutinase from Pseudomonas mendocina (U.S. Pat. No. 5,389,536), lipasefrom Thermobifida fusca (WO 11/084412), Geobacillus stearothermophiluslipase (WO 11/084417), lipase from Bacillus subtilis (WO 11/084599), andlipase from Streptomyces griseus (WO 11/150157) and S. pristinaespiralis(WO 12/137147).

Other examples are lipase variants such as those described in EP 407225,WO 92/05249, WO 94/01541, WO 94/25578, WO 95/14783, WO 95/30744, WO95/35381, WO 95/22615, WO 96/00292, WO 97/04079, WO 97/07202, WO00/34450, WO 00/60063, WO 01/92502, WO 07/87508 and WO 09/109500.

Preferred commercial lipase products include Lipolase™, Lipex™; Lipolex™and Lipoclean™ (Novozymes A/S), Lumafast (originally from Genencor) andLipomax (originally from Gist-Brocades).

Still other examples are lipases sometimes referred to asacyltransferases or perhydrolases, e.g. acyltransferases with homologyto Candida antarctica lipase A (WO 10/111143), acyltransferase fromMycobacterium smegmatis (WO 05/56782), perhydrolases from the CE 7family (WO 09/67279), and variants of the M. smegmatis perhydrolase inparticular the S54V variant used in the commercial product Gentle PowerBleach from Huntsman Textile Effects Pte Ltd (WO 10/100028).

Amylases

Suitable amylases which can be used together with the DNases of theinvention may be an alpha-amylase or a glucoamylase and may be ofbacterial or fungal origin. Chemically modified or protein engineeredmutants are included. Amylases include, for example, alpha-amylasesobtained from Bacillus, e.g., a special strain of Bacilluslicheniformis, described in more detail in GB 1,296,839.

Suitable amylases include amylases having SEQ ID NO: 2 in WO 95/10603 orvariants having 90% sequence identity to SEQ ID NO: 1 thereof. Preferredvariants are described in WO 94/02597, WO 94/18314, WO 97/43424 and SEQID NO: 4 of WO 99/019467, such as variants with substitutions in one ormore of the following positions: 15, 23, 105, 106, 124, 128, 133, 154,156, 178, 179, 181, 188, 190, 197, 201, 202, 207, 208, 209, 211, 243,264, 304, 305, 391, 408, and 444.

Different suitable amylases include amylases having SEQ ID NO: 6 in WO02/010355 or variants thereof having 90% sequence identity to SEQ ID NO:6. Preferred variants of SEQ ID NO: 6 are those having a deletion inpositions 181 and 182 and a substitution in position 193.

Other amylases which are suitable are hybrid alpha-amylase comprisingresidues 1-33 of the alpha-amylase obtained from B. amyloliquefaciensshown in SEQ ID NO: 6 of WO 2006/066594 and residues 36-483 of the B.licheniformis alpha-amylase shown in SEQ ID NO: 4 of WO 2006/066594 orvariants having 90% sequence identity thereof. Preferred variants ofthis hybrid alpha-amylase are those having a substitution, a deletion oran insertion in one of more of the following positions: G48, T49, G107,H156, A181, N190, M197, I201, A209 and Q264. Most preferred variants ofthe hybrid alpha-amylase comprising residues 1-33 of the alpha-amylaseobtained from B. amyloliquefaciens shown in SEQ ID NO: 6 of WO2006/066594 and residues 36-483 of SEQ ID NO: 4 are those having thesubstitutions:

M197T;

H156Y+A181T+N190F+A209V+Q264S; or

G48A+T491+G107A+H156Y+A181T+N190F+I201F+A209V+Q264S.

Further amylases which are suitable are amylases having SEQ ID NO: 6 inWO 99/019467 or variants thereof having 90% sequence identity to SEQ IDNO: 6. Preferred variants of SEQ ID NO: 6 are those having asubstitution, a deletion or an insertion in one or more of the followingpositions: R181, G182, H183, G184, N195, 1206, E212, E216 and K269.Particularly preferred amylases are those having deletion in positionsR181 and G182, or positions H183 and G184.

Additional amylases which can be used are those having SEQ ID NO: 1, SEQID NO: 3, SEQ ID NO: 2 or SEQ ID NO: 7 of WO 96/023873 or variantsthereof having 90% sequence identity to SEQ ID NO: 1, SEQ ID NO: 2, SEQID NO: 3 or SEQ ID NO: 7. Preferred variants of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO: 3 or SEQ ID NO: 7 are those having a substitution, adeletion or an insertion in one or more of the following positions: 140,181, 182, 183, 184, 195, 206, 212, 243, 260, 269, 304 and 476, using SEQID NO. 2 of WO 96/023873 for numbering. More preferred variants arethose having a deletion in two positions selected from 181, 182, 183 and184, such as 181 and 182, 182 and 183, or positions 183 and 184. Mostpreferred amylase variants of SEQ ID NO: 1, SEQ ID NO: 2 or SEQ ID NO: 7are those having a deletion in positions 183 and 184 and a substitutionin one or more of positions 140, 195, 206, 243, 260, 304 and 476.

Other amylases which can be used are amylases having SEQ ID NO: 2 of WO08/153815, SEQ ID NO: 10 in WO 01/66712 or variants thereof having 90%sequence identity to SEQ ID NO: 2 of WO 08/153815 or 90% sequenceidentity to SEQ ID NO: 10 in WO 01/66712. Preferred variants of SEQ IDNO: 10 in WO 01/66712 are those having a substitution, a deletion or aninsertion in one of more of the following positions: 176, 177, 178, 179,190, 201, 207, 211 and 264.

Further suitable amylases are amylases having SEQ ID NO: 2 of WO09/061380 or variants having 90% sequence identity to SEQ ID NO: 2thereof. Preferred variants of SEQ ID NO: 2 are those having atruncation of the C-terminus and/or a substitution, a deletion or aninsertion in one of more of the following positions: Q87, Q98, S125,N128, T131, T165, K178, R180, S181, T182, G183, M201, F202, N225, S243,N272, N282, Y305, R309, D319, Q320, Q359, K444 and G475. More preferredvariants of SEQ ID NO: 2 are those having the substitution in one ofmore of the following positions: Q87E,R, Q98R, S125A, N128C, T131I,T165I, K178L, T182G, M201L, F202Y, N225E,R, N272E,R, S243Q,A,E,D, Y305R,R309A, Q320R, Q359E, K444E and G475K and/or deletion in position R180and/or S181 or of T182 and/or G183. Most preferred amylase variants ofSEQ ID NO: 2 are those having the substitutions:

N128C+K178L+T182G+Y305R+G475K;

N128C+K178L+T182G+F202Y+Y305R+D319T+G475K;

S125A+N128C+K178L+T182G+Y305R+G475K; or

S125A+N128C+T1311+T165I+K178L+T182G+Y305R+G475K wherein the variants areC-terminally truncated and optionally further comprises a substitutionat position 243 and/or a deletion at position 180 and/or position 181.

Other suitable amylases are the alpha-amylase having SEQ ID NO: 12 in WO01/66712 or a variant having at least 90% sequence identity to SEQ IDNO: 12. Preferred amylase variants are those having a substitution, adeletion or an insertion in one of more of the following positions ofSEQ ID NO: 12 in WO01/66712: R28, R118, N174; R181, G182, D183, G184,G186, W189, N195, M202, Y298, N299, K302, S303, N306, R310, N314; R320,H324, E345, Y396, R400, W439, R444, N445, K446, Q449, R458, N471, N484.Particular preferred amylases include variants having a deletion of D183and G184 and having the substitutions R118K, N195F, R320K and R458K, anda variant additionally having substitutions in one or more positionselected from the group: M9, G149, G182, G186, M202, T257, Y295, N299,M323, E345 and A339, most preferred a variant that additionally hassubstitutions in all these positions.

Other examples are amylase variants such as those described in WO2011/098531, WO 2013/001078 and WO 2013/001087.

Commercially available amylases are Duramyl™, Termamyl™, Fungamyl™,Stainzyme™ Stainzyme Plus™, Natalase™, Liquozyme X and BAN™ (fromNovozymes A/S), and Rapidase™ Purastar™/Effectenz™, Powerase andPreferenz S100 (from Genencor International Inc./DuPont).

Peroxidases/Oxidases

A peroxidase according to the invention is a peroxidase enzyme comprisedby the enzyme classification EC 1.11.1.7, as set out by the NomenclatureCommittee of the International Union of Biochemistry and MolecularBiology (IUBMB), or any fragment obtained therefrom, exhibitingperoxidase activity.

Suitable peroxidases include those of plant, bacterial or fungal origin.Chemically modified or protein engineered mutants are included. Examplesof useful peroxidases include peroxidases from Coprinopsis, e.g., fromC. cinerea (EP 179,486), and variants thereof as those described in WO93/24618, WO 95/10602, and WO 98/15257.

A peroxidase according to the invention also includes a haloperoxidaseenzyme, such as chloroperoxidase, bromoperoxidase and compoundsexhibiting chloroperoxidase or bromoperoxidase activity. Haloperoxidasesare classified according to their specificity for halide ions.Chloroperoxidases (E.C. 1.11.1.10) catalyze formation of hypochloritefrom chloride ions.

In an embodiment, the haloperoxidase of the invention is achloroperoxidase. Preferably, the haloperoxidase is a vanadiumhaloperoxidase, i.e., a vanadate-containing haloperoxidase. In apreferred method of the present invention the vanadate-containinghaloperoxidase is combined with a source of chloride ion.

Haloperoxidases have been isolated from many different fungi, inparticular from the fungus group dematiaceous hyphomycetes, such asCaldariomyces, e.g., C. fumago, Alternaria, Curvularia, e.g., C.verruculosa and C. inaequalis, Drechslera, Ulocladium and Botrytis.

Haloperoxidases have also been isolated from bacteria such asPseudomonas, e.g., P. pyrrocinia and Streptomyces, e.g., S.aureofaciens.

In a preferred embodiment, the haloperoxidase is derivable fromCurvularia sp., in particular Curvularia verruculosa or Curvulariainaequalis, such as C. inaequalis CBS 102.42 as described in WO95/27046; or C. verruculosa CBS 147.63 or C. verruculosa CBS 444.70 asdescribed in WO 97/04102; or from Drechslera hartlebii as described inWO 01/79459, Dendryphiella salina as described in WO 01/79458,Phaeotrichoconis crotalarie as described in WO 01/79461, orGeniculosporium sp. as described in WO 01/79460.

An oxidase according to the invention include, in particular, anylaccase enzyme comprised by the enzyme classification EC 1.10.3.2, orany fragment obtained therefrom exhibiting laccase activity, or acompound exhibiting a similar activity, such as a catechol oxidase (EC1.10.3.1), an o-aminophenol oxidase (EC 1.10.3.4), or a bilirubinoxidase (EC 1.3.3.5).

Preferred laccase enzymes are enzymes of microbial origin. The enzymesmay be obtained from plants, bacteria or fungi (including filamentousfungi and yeasts).

Suitable examples from fungi include a laccase derivable from a strainof Aspergillus, Neurospora, e.g., N. crassa, Podospora, Botrytis,Collybia, Fomes, Lentinus, Pleurotus, Trametes, e.g., T. villosa and T.versicolor, Rhizoctonia, e.g., R. solani, Coprinopsis, e.g., C. cinerea,C. comatus, C. friesii, and C. plicatilis, Psathyrella, e.g., P.condelleana, Panaeolus, e.g., P. papilionaceus, Myceliophthora, e.g., M.thermophila, Schytalidium, e.g., S. thermophilum, Polyporus, e.g., P.pinsitus, Phlebia, e.g., P. radiata (WO 92/01046), or Coriolus, e.g., C.hirsutus (JP 2238885).

Suitable examples from bacteria include a laccase derivable from astrain of Bacillus.

A laccase obtained from Coprinopsis or Myceliophthora is preferred; inparticular a laccase obtained from Coprinopsis cinerea, as disclosed inWO 97/08325; or from Myceliophthora thermophila, as disclosed in WO95/33836.

The detergent enzyme(s) may be included in a detergent composition byadding separate additives containing one or more enzymes, or by adding acombined additive comprising all of these enzymes. A detergent additiveof the invention, i.e., a separate additive or a combined additive, canbe formulated, for example, as a granulate, liquid, slurry, etc.Preferred detergent additive formulations are granulates, in particularnon-dusting granulates, liquids, in particular stabilized liquids, orslurries.

Non-dusting granulates may be produced, e.g. as disclosed in U.S. Pat.Nos. 4,106,991 and 4,661,452 and may optionally be coated by methodsknown in the art. Examples of waxy coating materials are poly(ethyleneoxide) products (polyethyleneglycol, PEG) with mean molar weights of1000 to 20000; ethoxylated nonylphenols having from 16 to 50 ethyleneoxide units; ethoxylated fatty alcohols in which the alcohol containsfrom 12 to 20 carbon atoms and in which there are 15 to 80 ethyleneoxide units; fatty alcohols; fatty acids; and mono- and di- andtriglycerides of fatty acids. Examples of film-forming coating materialssuitable for application by fluid bed techniques are given in GB1483591. Liquid enzyme preparations may, for instance, be stabilized byadding a polyol such as propylene glycol, a sugar or sugar alcohol,lactic acid or boric acid according to established methods. Protectedenzymes may be prepared according to the method disclosed in EP 238,216.

Other Materials

Any detergent components known in the art for use in the cleaningcomposition of the invention may also be utilized. Other optionaldetergent components include anti-corrosion agents, anti-shrink agents,anti-soil redeposition agents, anti-wrinkling agents, bactericides,binders, corrosion inhibitors, disintegrants/disintegration agents,dyes, enzyme stabilizers (including boric acid, borates, CMC, and/orpolyols such as propylene glycol), fabric conditioners including clays,fillers/processing aids, fluorescent whitening agents/opticalbrighteners, foam boosters, foam (suds) regulators, perfumes,soil-suspending agents, softeners, suds suppressors, tarnish inhibitors,and wicking agents, either alone or in combination. Any ingredient knownin the art for use in detergents may be utilized. The choice of suchingredients is well within the skill of the artisan.

Dispersants

The detergent compositions of the present invention can also containdispersants. In particular powdered detergents may comprise dispersants.Suitable water-soluble organic materials include the homo- orco-polymeric acids or their salts, in which the polycarboxylic acidcomprises at least two carboxyl radicals separated from each other bynot more than two carbon atoms. Suitable dispersants are for exampledescribed in Powdered Detergents, Surfactant science series volume 71,Marcel Dekker, Inc.

Dye Transfer Inhibiting Agents

The cleaning compositions of the present invention may also include oneor more dye transfer inhibiting agents. Suitable polymeric dye transferinhibiting agents include, but are not limited to, polyvinylpyrrolidonepolymers, polyamine N-oxide polymers, copolymers of N-vinylpyrrolidoneand N-vinylimidazole, polyvinyloxazolidones and polyvinylimidazoles ormixtures thereof. When present in a subject composition, the dyetransfer inhibiting agents may be present at levels from about 0.0001%to about 10%, from about 0.01% to about 5% or even from about 0.1% toabout 3% by weight of the composition.

Fluorescent Whitening Agent

The detergent composition may preferably also contain additionalcomponents that may tint articles being cleaned, such as fluorescentwhitening agent or optical brighteners. Where present the brightener ispreferably at a level of about 0.01% to about 0.5%. Any fluorescentwhitening agent suitable for use in a laundry detergent composition maybe used in the composition of the present invention. The most commonlyused fluorescent whitening agents are those belonging to the classes ofdiaminostilbene-sulfonic acid derivatives, diarylpyrazoline derivativesand bisphenyl-distyryl derivatives. Examples of thediaminostilbene-sulfonic acid derivative type of fluorescent whiteningagents include the sodium salts of:4,4′-bis-(2-diethanolamino-4-anilino-s-triazin-6-ylamino)stilbene-2,2′-disulfonate, 4,4′-bis-(2,4-dianilino-s-triazin-6-ylamino)stilbene-2.2′-disulfonate,4,4′-bis-(2-anilino-4-(N-methyl-N-2-hydroxy-ethylamino)-s-triazin-6-ylamino)stilbene-2,2′-disulfonate,4,4′-bis-(4-phenyl-1,2,3-triazol-2-yl)stilbene-2,2′-disulfonate andsodium5-(2H-naphtho[1,2-d][1,2,3]triazol-2-yl)-2-[(E)-2-phenylvinyl]benzenesulfonate.Preferred fluorescent whitening agents are Tinopal DMS and Tinopal CBSavailable from Ciba-Geigy AG, Basel, Switzerland. Tinopal DMS is thedisodium salt of 4,4′-bis-(2-morpholino-4-anilino-s-triazin-6-ylamino)stilbene-2,2′-disulfonate. Tinopal CBS is the disodium salt of2,2′-bis-(phenyl-styryl)-disulfonate. Also preferred are fluorescentwhitening agents is the commercially available Parawhite KX, supplied byParamount Minerals and Chemicals, Mumbai, India. Tinopal CBS-X is a4.4′-bis-(sulfostyryl)-biphenyl disodium salt also known as DisodiumDistyrylbiphenyl Disulfonate. Other fluorescers suitable for use in theinvention include the 1-3-diaryl pyrazolines and the7-alkylaminocoumarins.

Suitable fluorescent brightener levels include lower levels of fromabout 0.01, from 0.05, from about 0.1 or even from about 0.2 wt % toupper levels of 0.5 or even 0.75 wt %.

Soil Release Polymers

The detergent compositions may also include one or more soil releasepolymers which aid the removal of soils from fabrics such as cotton andpolyester based fabrics, in particular the removal of hydrophobic soilsfrom polyester based fabrics. The soil release polymers may for examplebe nonionic or anionic terephthalte based polymers, polyvinylcaprolactam and related copolymers, vinyl graft copolymers, polyesterpolyamides see for example Chapter 7 in Powdered Detergents, Surfactantscience series volume 71, Marcel Dekker, Inc. Another type of soilrelease polymers are amphiphilic alkoxylated grease cleaning polymerscomprising a core structure and a plurality of alkoxylate groupsattached to that core structure. The core structure may comprise apolyalkylenimine structure or a polyalkanolamine structure as describedin detail in WO 2009/087523 (hereby incorporated by reference).Furthermore random graft co-polymers are suitable soil release polymers.Suitable graft co-polymers are described in more detail in WO2007/138054, WO 2006/108856 and WO 2006/113314 (hereby incorporated byreference). Other soil release polymers are substituted polysaccharidestructures especially substituted cellulosic structures such as modifiedcellulose deriviatives such as those described in EP 1867808 or WO2003/040279 (both are hereby incorporated by reference). Suitablecellulosic polymers include cellulose, cellulose ethers, celluloseesters, cellulose amides and mixtures thereof. Suitable cellulosicpolymers include anionically modified cellulose, nonionically modifiedcellulose, cationically modified cellulose, zwitterionically modifiedcellulose, and mixtures thereof. Suitable cellulosic polymers includemethyl cellulose, carboxy methyl cellulose, ethyl cellulose, hydroxylethyl cellulose, hydroxyl propyl methyl cellulose, ester carboxy methylcellulose, and mixtures thereof.

Anti-Redeposition Agents

The detergent compositions of the present invention may also include oneor more anti-redeposition agents such as carboxymethylcellulose (CMC),polyvinyl alcohol (PVA), polyvinylpyrrolidone (PVP), polyoxyethyleneand/or polyethyleneglycol (PEG), homopolymers of acrylic acid,copolymers of acrylic acid and maleic acid, and ethoxylatedpolyethyleneimines. The cellulose based polymers described under soilrelease polymers above may also function as anti-redeposition agents.

Rheology Modifiers

The detergent compositions of the present invention may also include oneor more rheology modifiers, structurants or thickeners, as distinct fromviscosity reducing agents. The rheology modifiers are selected from thegroup consisting of non-polymeric crystalline, hydroxy-functionalmaterials, polymeric rheology modifiers which impart shear thinningcharacteristics to the aqueous liquid matrix of a liquid detergentcomposition. The rheology and viscosity of the detergent can be modifiedand adjusted by methods known in the art, for example as shown in EP2169040.

Other suitable adjunct materials include, but are not limited to,anti-shrink agents, anti-wrinkling agents, bactericides, binders,carriers, dyes, enzyme stabilizers, fabric softeners, fillers, foamregulators, hydrotropes, perfumes, pigments, sod suppressors, solvents,and structurants for liquid detergents and/or structure elasticizingagents.

Formulation of Detergent Products

The detergent composition may be in any convenient form, e.g., a bar, ahomogenous tablet, a tablet having two or more layers, a regular orcompact powder, a granule, a paste, a gel, or a regular, compact orconcentrated liquid.

Detergent formulation forms: Layers (same or different phases), Pouches,versus forms for machine dosing unit.

Pouches can be configured as single or multicompartments. It can be ofany form, shape and material which is suitable for hold the composition,e.g. without allowing the release of the composition to release of thecomposition from the pouch prior to water contact. The pouch is madefrom water soluble film which encloses an inner volume. Said innervolume can be divided into compartments of the pouch. Preferred filmsare polymeric materials preferably polymers which are formed into a filmor sheet. Preferred polymers, copolymers or derivates thereof areselected polyacrylates, and water soluble acrylate copolymers, methylcellulose, carboxy methyl cellulose, sodium dextrin, ethyl cellulose,hydroxyethyl cellulose, hydroxypropyl methyl cellulose, malto dextrin,poly methacrylates, most preferably polyvinyl alcohol copolymers and,hydroxyprpyl methyl cellulose (HPMC). Preferably the level of polymer inthe film for example PVA is at least about 60%. Preferred averagemolecular weight will typically be about 20,000 to about 150,000. Filmscan also be of blend compositions comprising hydrolytically degradableand water soluble polymer blends such as polyactide and polyvinylalcohol (known under the Trade reference M8630 as sold by Chris CraftIn. Prod. Of Gary, Ind., US) plus plasticisers like glycerol, ethyleneglycerol, Propylene glycol, sorbitol and mixtures thereof. The pouchescan comprise a solid laundry cleaning composition or part componentsand/or a liquid cleaning composition or part components separated by thewater soluble film. The compartment for liquid components can bedifferent in composition than compartments containing solids. Ref: (US2009/0011970 A1)

Detergent ingredients can be separated physically from each other bycompartments in water dissolvable pouches or in different layers oftablets. Thereby negative storage interaction between components can beavoided. Different dissolution profiles of each of the compartments canalso give rise to delayed dissolution of selected components in the washsolution.

Definition/Characteristics of the Forms:

A liquid or gel detergent, which is not unit dosed, may be aqueous,typically containing at least 20% by weight and up to 95% water, such asup to about 70% water, up to about 65% water, up to about 55% water, upto about 45% water, up to about 35% water. Other types of liquids,including without limitation, alkanols, amines, diols, ethers andpolyols may be included in an aqueous liquid or gel. An aqueous liquidor gel detergent may contain from 0-30% organic solvent.

A liquid or gel detergent may be non-aqueous.

Granular Detergent Formulations

A granular detergent may be formulated as described in WO 09/092699, EP1705241, EP 1382668, WO 07/001262, U.S. Pat. No. 6,472,364, WO 04/074419or WO 09/102854. Other useful detergent formulations are described in WO09/124162, WO 09/124163, WO 09/117340, WO 09/117341, WO 09/117342, WO09/072069, WO 09/063355, WO 09/132870, WO 09/121757, WO 09/112296, WO09/112298, WO 09/103822, WO 09/087033, WO 09/050026, WO 09/047125, WO09/047126, WO 09/047127, WO 09/047128, WO 09/021784, WO 09/010375, WO09/000605, WO 09/122125, WO 09/095645, WO 09/040544, WO 09/040545, WO09/024780, WO 09/004295, WO 09/004294, WO 09/121725, WO 09/115391, WO09/115392, WO 09/074398, WO 09/074403, WO 09/068501, WO 09/065770, WO09/021813, WO 09/030632, WO 09/015951, WO 2011025615, WO 2011016958, WO2011005803, WO 2011005623, WO 2011005730, WO 2011005844, WO 2011005904,WO 2011005630, WO 2011005830, WO 2011005912, WO 2011005905, WO2011005910, WO 2011005813, WO 2010135238, WO 2010120863, WO 2010108002,WO 2010111365, WO 2010108000, WO 2010107635, WO 2010090915, WO2010033976, WO 2010033746, WO 2010033747, WO 2010033897, WO 2010033979,WO 2010030540, WO 2010030541, WO 2010030539, WO 2010024467, WO2010024469, WO 2010024470, WO 2010025161, WO 2010014395, WO 2010044905,WO 2010145887, WO 2010142503, WO 2010122051, WO 2010102861, WO2010099997, WO 2010084039, WO 2010076292, WO 2010069742, WO 2010069718,WO 2010069957, WO 2010057784, WO 2010054986, WO 2010018043, WO2010003783, WO 2010003792, WO 2011023716, WO 2010142539, WO 2010118959,WO 2010115813, WO 2010105942, WO 2010105961, WO 2010105962, WO2010094356, WO 2010084203, WO 2010078979, WO 2010072456, WO 2010069905,WO 2010076165, WO 2010072603, WO 2010066486, WO 2010066631, WO2010066632, WO 2010063689, WO 2010060821, WO 2010049187, WO 2010031607,WO 2010000636.

Formulation of Enzyme in Co-Granule

The DNase may be formulated as a granule for example as a co-granulethat combines one or more enzymes. Each enzyme will then be present inmore granules securing a more uniform distribution of enzymes in thedetergent. This also reduces the physical segregation of differentenzymes due to different particle sizes. Methods for producingmulti-enzyme co-granulate for the detergent industry is disclosed in theIP.com disclosure IPCOM000200739D.

Another example of formulation of enzymes by the use of co-granulatesare disclosed in WO 2013/188331, which relates to a detergentcomposition comprising (a) a multi-enzyme co-granule; (b) less than 10wt zeolite (anhydrous basis); and (c) less than 10 wt phosphate salt(anhydrous basis), wherein said enzyme co-granule comprises from 10 to98 wt % moisture sink component and the composition additionallycomprises from 20 to 80 wt % detergent moisture sink component. WO2013/188331 also relates to a method of treating and/or cleaning asurface, preferably a fabric surface comprising the steps of (i)contacting said surface with the detergent composition as claimed anddescribed herein in aqueous wash liquor, (ii) rinsing and/or drying thesurface.

The invention is further described in the following non-limitingembodiments;

1. A DNase variant, comprising an alteration compared to SEQ ID NO: 1 atone or more positions corresponding to positions 1, 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26,27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44,45, 46, 47, 48, 49, 50, 51, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63,64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81,82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99,100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113,114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127,128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141,142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155,156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169,170, 171, 172, 173, 174, 175, 176, 177, 178, 179, 180, 181, 182, 183,185, 186, 187, 189, 190, 191, 192, 193, 194, 195, 196, 197, 198, 199,200, 201, 202, 203, 204, 205, 206, 207, 208, 209, 210, 211, 212, 213,214, 215, 216, 217, 218, 219, 220 or 221 of SEQ ID NO: 1, wherein thevariant has a sequence identity to SEQ ID NO: 1 of at least 60% and thevariant has DNase activity.2. A variant of embodiment 1, which comprises one or more alterationscompared to SEQ ID NO: 1, wherein the one or more alteration(s) isselected from the group consisting of V1*, V1A, V1D, V1E, V1F, V1G, V1H,V1I, V1K, V1L, V1M, V1N, V1P, V1Q, V1R, V1S, V1T, V1W, V1Y, P2*, P2A,P2D, P2E, P2F, P2G, P2H, P2I, P2K, P2L, P2M, P2N, P2Q, P2R, P2S, P2T,P2V, P2W, P2Y, V3*, V3A, V3C, V3D, V3E, V3F, V3G, V3H, V3I, V3K, V3L,V3M, V3N, V3P, V3R, V3S, V3T, V3W, V3Y, N4*, N4A, N4D, N4E, N4F, N4G,N4H, N4I, N4K, N4L, N4M, N4P, N4Q, N4R, N4S, N4T, N4V, N4W, N4Y, P5*,P5A, P5D, P5E, P5F, P5G, P5H, P5I, P5K, P5L, P5M, P5N, P5Q, P5R, P5S,P5T, P5V, P5W, P5Y, E6*, E6A, E6D, E6F, E6G, E6H, E6I, E6K, E6L, E6M,E6N, E6P, E6Q, E6R, E6S, E6T, E6V, E6W, E6Y, P7*, P7A, P7D, P7E, P7F,P7G, P7H, P7I, P7K, P7L, P7M, P7N, P7Q, P7R, P7S, P7T, P7V, P7W, P7Y,D8*, D8A, D8E, D8F, D8G, D8H, D8I, D8K, D8L, D8M, D8N, D8P, D8Q, D8R,D8S, D8T, D8V, D8W, D8Y, A9*, A9D, A9E, A9F, A9G, A9H, A9I, A9K, A9L,A9M, A9N, A9P, A9Q, A9R, A9S, A9T, A9V, A9W, A9Y, T10*, T10A, T10D,T10E, T10F, T10G, T10H, T10I, T10K, T10L, T10M, T10N, T10P, T10Q, T10R,T10S, T10V, T10W, T10Y, S11*, S11A, S11D, S11E, S11F, S11G, S11H, S11I,S11K, S11L, S11M, S11N, S11P, S11Q, S11R, S11T, S11V, S11W, S11Y, V12*,V12A, V12D, V12E, V12F, V12G, V12H, V12I, V12K, V12L, V12M, V12N, V12P,V12Q, V12R, V12S, V12T, V12W, V12Y, E13*, E13A, E13D, E13F, E13G, E13H,E13I, E13K, E13L, E13M, E13N, E13P, E13Q, E13R, E13S, E13T, E13V, E13W,E13Y, N14*, N14A, N14D, N14E, N14F, N14G, N14H, N14I, N14K, N14L, N14M,N14P, N14Q, N14R, N14S, N14T, N14V, N14W, N14Y, V15*, V15A, V15D, V15E,V15F, V15G, V15H, V15I, V15K, V15L, V15M, V15N, V15P, V15Q, V15R, V15S,V15T, V15W, V15Y, A16*, A16D, A16E, A16F, A16G, A16H, A16I, A16K, A16L,A16M, A16N, A16P, A16Q, A16R, A16S, A16T, A16V, A16W, A16Y, L17*, L17A,L17D, L17E, L17F, L17G, L17H, L17I, L17K, L17M, L17N, L17P, L17Q, L17R,L17S, L17T, L17V, L17W, L17Y, K18*, K18A, K18D, K18E, K18F, K18G, K18H,K18I, K18L, K18M, K18N, K18P, K18Q, K18R, K18S, K18T, K18V, K18W, K18Y,T19*, T19A, T19D, T19E, T19F, T19G, T19H, T19I, T19K, T19L, T19M, T19N,T19P, T19Q, T19R, T19S, T19V, T19W, T19Y, G20*, G20A, G20D, G20E, G20F,G20H, G20I, G20K, G20L, G20M, G20N, G20P, G20Q, G20R, G20S, G20T, G20V,G20W, G20Y, S21*, S21A, S21D, S21E, S21F, S21G, S21H, S21I, S21K, S21L,S21M, S21N, S21P, S21Q, S21R, S21T, S21V, S21W, S21Y, G22*, G22A, G22D,G22E, G22F, G22H, G22I, G22K, G22L, G22M, G22N, G22P, G22Q, G22R, G22S,G22T, G22V, G22W, G22Y, D23*, D23A, D23E, D23F, D23G, D23H, D23I, D23K,D23L, D23M, D23N, D23P, D23Q, D23R, D23S, D23T, D23V, D23W, D23Y, S24*,S24A, S24D, S24E, S24F, S24G, S24H, S24I, S24K, S24L, S24M, S24N, S24P,S24Q, S24R, S24T, S24V, S24W, S24Y, Q25*, Q25A, Q25D, Q25E, Q25F, Q25G,Q25H, Q25I, Q25K, Q25L, Q25M, Q25N, Q25P, Q25R, Q25S, Q25T, Q25V, Q25W,Q25Y, S26*, S26A, S26D, S26E, S26F, S26G, S26H, S26I, S26K, S26L, S26M,S26N, S26P, S26Q, S26R, S26T, S26V, S26W, S26Y, D27*, D27A, D27E, D27F,D27G, D27H, D27I, D27K, D27L, D27M, D27N, D27P, D27Q, D27R, D27S, D27T,D27V, D27W, D27Y, P28*, P28A, P28D, P28E, P28F, P28G, P28H, P28I, P28K,P28L, P28M, P28N, P28Q, P28R, P28S, P28T, P28V, P28W, P28Y, I29*, I29A,I29D, I29E, I29F, I29G, I29H, I29K, I29L, I29M, I29N, I29P, I29Q, I29R,I29S, I29T, I29V, I29W, I29Y, K30*, K30A, K30D, K30E, K30F, K30G, K30H,K30I, K30L, K30M, K30N, K30P, K30Q, K30R, K30S, K30T, K30V, K30W, K30Y,A31*, A31D, A31E, A31F, A31G, A31H, A31I, A31K, A31L, A31M, A31N, A31P,A31Q, A31R, A31S, A31T, A31V, A31W, A31Y, D32*, D32A, D32E, D32F, D32G,D32H, D32I, D32K, D32L, D32M, D32N, D32P, D32Q, D32R, D32S, D32T, D32V,D32W, D32Y, L33*, L33A, L33D, L33E, L33F, L33G, L33H, L33I, L33K, L33M,L33N, L33P, L33Q, L33R, L33S, L33T, L33V, L33W, L33Y, E34*, E34A, E34D,E34F, E34G, E34H, E34I, E34K, E34L, E34M, E34N, E34P, E34Q, E34R, E34S,E34T, E34V, E34W, E34Y, V35*, V35A, V35D, V35E, V35F, V35G, V35H, V35I,V35K, V35L, V35M, V35N, V35P, V35Q, V35R, V35S, V35T, V35W, V35Y, K36*,K36A, K36D, K36E, K36F, K36G, K36H, K36I, K36L, K36M, K36N, K36P, K36Q,K36R, K36S, K36T, K36V, K36W, K36Y, G37*, G37A, G37D, G37E, G37F, G37H,G37I, G37K, G37L, G37M, G37N, G37P, G37Q, G37R, G37S, G37T, G37V, G37W,G37Y, Q38*, Q38A, Q38D, Q38E, Q38F, Q38G, Q38H, Q38I, Q38K, Q38L, Q38M,Q38N, Q38P, Q38R, Q38S, Q38T, Q38V, Q38W, Q38Y, S39*, S39A, S39D, S39E,S39F, S39G, S39H, S39I, S39K, S39L, S39M, S39N, S39P, S39Q, S39R, S39T,S39V, S39W, S39Y, A40*, A40D, A40E, A40F, A40G, A40H, A40I, A40K, A40L,A40M, A40N, A40P, A40Q, A40R, A40S, A40T, A40V, A40W, A40Y, L41*, L41A,L41D, L41E, L41F, L41G, L41H, L41I, L41K, L41M, L41N, L41P, L41Q, L41R,L41S, L41T, L41V, L41W, L41Y, P42*, P42A, P42D, P42E, P42F, P42G, P42H,P42I, P42K, P42L, P42M, P42N, P42Q, P42R, P42S, P42T, P42V, P42W, P42Y,F43*, F43A, F43D, F43E, F43G, F43H, F43I, F43K, F43L, F43M, F43N, F43P,F43Q, F43R, F43S, F43T, F43V, F43W, F43Y, D44*, D44A, D44E, D44F, D44G,D44H, D44I, D44K, D44L, D44M, D44N, D44P, D44Q, D44R, D44S, D44T, D44V,D44W, D44Y, V45*, V45A, V45D, V45E, V45F, V45G, V45H, V45I, V45K, V45L,V45M, V45N, V45P, V45Q, V45R, V45S, V45T, V45W, V45Y, D46*, D46A, D46E,D46F, D46G, D46H, D46I, D46K, D46L, D46M, D46N, D46P, D46Q, D46R, D46S,D46T, D46V, D46W, D46Y, C47A, C47D, C47E, C47F, C47G, C47H, C47I, C47K,C47L, C47M, C47N, C47P, C47Q, C47R, C47S, C47T, C47V, C47W, C47Y, W48*,W48A, W48D, W48E, W48F, W48G, W48H, W48I, W48K, W48L, W48M, W48N, W48P,W48Q, W48R, W48S, W48T, W48V, W48Y, A49*, A49D, A49E, A49F, A49G, A49H,A49I, A49K, A49L, A49M, A49N, A49P, A49Q, A49R, A49S, A49T, A49V, A49W,A49Y, I50*, I50A, I50D, I50E, I50F, I50G, I50H, I50K, I50L, I50M, I50N,I50P, I50Q, I50R, I50S, I50T, I50V, I50W, I50Y, L51*, L51A, L51D, L51E,L51F, L51G, L51H, L51I, L51K, L51M, L51N, L51P, L51Q, L51R, L51S, L51T,L51V, L51W, L51Y, K53*, K53A, K53D, K53E, K53F, K53G, K53H, K53I, K53L,K53M, K53N, K53P, K53Q, K53R, K53S, K53T, K53V, K53W, K53Y, G54*, G54A,G54D, G54E, G54F, G54H, G54I, G54K, G54L, G54M, G54N, G54P, G54Q, G54R,G54S, G54T, G54V, G54W, G54Y, A55*, A55D, A55E, A55F, A55G, A55H, A55I,A55K, A55L, A55M, A55N, A55P, A55Q, A55R, A55S, A55T, A55V, A55W, A55Y,P56*, P56A, P56D, P56E, P56F, P56G, P56H, P56I, P56K, P56L, P56M, P56N,P56Q, P56R, P56S, P56T, P56V, P56W, P56Y, N57*, N57A, N57D, N57E, N57F,N57G, N57H, N57I, N57K, N57L, N57M, N57P, N57Q, N57R, N57S, N57T, N57V,N57W, N57Y, V58*, V58A, V58D, V58E, V58F, V58G, V58H, V58I, V58K, V58L,V58M, V58N, V58P, V58Q, V58R, V58S, V58T, V58W, V58YL59*, L59A, L59A,L59D, L59D, L59A, L59D, L59E, L59E, L59F, L59F, L59E, L59F, L59G, L59G,L59H, L59H, L59G, L59H, L59I, L59I, L59K, L59K, L59I, L59K, L59M,L59ML59M, L59N, L59N, L59N, L59P, L59P, L59Q, L59Q, L59P, L59Q, L59R,L59R, L59S, L59S, L59R, L59S, L59T, L59T, L59V, L59V, L59T, L59V, L59W,L59W, L59Y, L59YL59W, L59Y, Q60*, Q60A, Q60D, Q60E, Q60F, Q60G, Q60H,Q60I, Q60K, Q60L, Q60M, Q60N, Q60P, Q60R, Q60S, Q60T, Q60V, Q60W, Q60Y,R61*, R61A, R61D, R61E, R61F, R61G, R61H, R61I, R61K, R61L, R61M, R61N,R61P, R61Q, R61S, R61T, R61V, R61W, R61Y, V62*, V62A, V62D, V62E, V62F,V62G, V62H, V62I, V62K, V62L, V62M, V62N, V62P, V62Q, V62R, V62S, V62T,V62W, V62Y, N63*, N63A, N63D, N63E, N63F, N63G, N63H, N63I, N63K, N63L,N63M, N63P, N63Q, N63R, N63S, N63T, N63V, N63W, N63Y, E64*, E64A, E64D,E64F, E64G, E64H, E64I, E64K, E64L, E64M, E64N, E64P, E64Q, E64R, E64S,E64T, E64V, E64W, E64Y, K65*, K65A, K65D, K65E, K65F, K65G, K65H, K65I,K65L, K65M, K65N, K65P, K65Q, K65R, K65S, K65T, K65V, K65W, K65Y, T66*,T66A, T66D, T66E, T66F, T66G, T66H, T66I, T66K, T66L, T66M, T66N, T66P,T66Q, T66R, T66S, T66V, T66W, T66Y, K67*, K67A, K67D, K67E, K67F, K67G,K67H, K67I, K67L, K67M, K67N, K67P, K67Q, K67R, K67S, K67T, K67V, K67W,K67Y, N68*, N68A, N68D, N68E, N68F, N68G, N68H, N68I, N68K, N68L, N68M,N68P, N68Q, N68R, N68S, N68T, N68V, N68W, N68Y, S69*, S69A, S69D, S69E,S69F, S69G, S69H, S69I, S69K, S69L, S69M, S69N, S69P, S69Q, S69R, S69T,S69V, S69W, S69Y, N70*, N70A, N70D, N70E, N70F, N70G, N70H, N70I, N70K,N70L, N70M, N70P, N70Q, N70R, N70S, N70T, N70V, N70W, N70Y, R71*, R71A,R71D, R71E, R71F, R71G, R71H, R71I, R71K, R71L, R71M, R71N, R71P, R71Q,R71S, R71T, R71V, R71W, R71Y, D72*, D72A, D72E, D72F, D72G, D72H, D72I,D72K, D72L, D72M, D72N, D72P, D72Q, D72R, D72S, D72T, D72V, D72W, D72Y,R73*, R73A, R73D, R73E, R73F, R73G, R73H, R73I, R73K, R73L, R73M, R73N,R73P, R73Q, R73S, R73T, R73V, R73W, R73Y, S74*, S74A, S74D, S74E, S74F,S74G, S74H, S74I, S74K, S74L, S74M, S74N, S74P, S74Q, S74R, S74T, S74V,S74W, S74Y, G75*, G75A, G75D, G75E, G75F, G75H, G75I, G75K, G75L, G75M,G75N, G75P, G75Q, G75R, G75S, G75T, G75V, G75W, G75Y, A76*, A76D, A76E,A76F, A76G, A76H, A76I, A76K, A76L, A76M, A76N, A76P, A76Q, A76R, A76S,A76T, A76V, A76W, A76Y, N77*, N77A, N77D, N77E, N77F, N77G, N77H, N77I,N77K, N77L, N77M, N77P, N77Q, N77R, N77S, N77T, N77V, N77W, N77Y, K78*,K78A, K78D, K78E, K78F, K78G, K78H, K78I, K78L, K78M, K78N, K78P, K78Q,K78R, K78S, K78T, K78V, K78W, K78Y, G79*, G79A, G79D, G79E, G79F, G79H,G79I, G79K, G79L, G79M, G79N, G79P, G79Q, G79R, G79S, G79T, G79V, G79W,G79Y, P80*, P80A, P80D, P80E, P80F, P80G, P80H, P80I, P80K, P80L, P80M,P80N, P80Q, P80R, P80S, P80T, P80V, P80W, P80Y, F81*, F81A, F81D, F81E,F81G, F81H, F81I, F81K, F81L, F81M, F81N, F81P, F81Q, F81R, F81S, F81T,F81V, F81W, F81Y, K82*, K82A, K82D, K82E, K82F, K82G, K82H, K82I, K82L,K82M, K82N, K82P, K82Q, K82R, K82S, K82T, K82V, K82W, K82Y, D83*, D83A,D83E, D83F, D83G, D83H, D83I, D83K, D83L, D83M, D83N, D83P, D83Q, D83R,D83S, D83T, D83V, D83W, D83Y, P84*, P84A, P84D, P84E, P84F, P84G, P84H,P84I, P84K, P84L, P84M, P84N, P84Q, P84R, P84S, P84T, P84V, P84W, P84Y,Q85*, Q85A, Q85D, Q85E, Q85F, Q85G, Q85H, Q85I, Q85K, Q85L, Q85M, Q85N,Q85P, Q85R, Q85S, Q85T, Q85V, Q85W, Q85Y, K86*, K86A, K86D, K86E, K86F,K86G, K86H, K86I, K86L, K86M, K86N, K86P, K86Q, K86R, K86S, K86T, K86V,K86W, K86Y, W87*, W87A, W87D, W87E, W87F, W87G, W87H, W87I, W87K, W87L,W87M, W87N, W87P, W87Q, W87R, W87S, W87T, W87V, W87Y, G88*, G88A, G88D,G88E, G88F, G88H, G88I, G88K, G88L, G88M, G88N, G88P, G88Q, G88R, G88S,G88T, G88V, G88W, G88Y, I89*, I89A, I89D, I89E, I89F, I89G, I89H, I89K,I89L, I89M, I89N, I89P, I89Q, I89R, I89S, I89T, I89V, I89W, I89Y, K90*,K90A, K90D, K90E, K90F, K90G, K90H, K90I, K90L, K90M, K90N, K90P, K90Q,K90R, K90S, K90T, K90V, K90W, K90Y, A91*, A91D, A91E, A91F, A91G, A91H,A91I, A91K, A91L, A91M, A91N, A91P, A91Q, A91R, A91S, A91T, A91V, A91W,A91Y, L92*, L92A, L92D, L92E, L92F, L92G, L92H, L92I, L92K, L92M, L92N,L92P, L92Q, L92R, L92S, L92T, L92V, L92W, L92Y, P93*, P93A, P93D, P93E,P93F, P93G, P93H, P93I, P93K, P93L, P93M, P93N, P93Q, P93R, P93S, P93T,P93V, P93W, P93Y, P94*, P94A, P94D, P94E, P94F, P94G, P94H, P94I, P94K,P94L, P94M, P94N, P94Q, P94R, P94S, P94T, P94V, P94W, P94Y, K95*, K95A,K95D, K95E, K95F, K95G, K95H, K95I, K95L, K95M, K95N, K95P, K95Q, K95R,K95S, K95T, K95V, K95W, K95Y, N96*, N96A, N96D, N96E, N96F, N96G, N96H,N96I, N96K, N96L, N96M, N96P, N96Q, N96R, N96S, N96T, N96V, N96W, N96Y,P97*, P97A, P97D, P97E, P97F, P97G, P97H, P97I, P97K, P97L, P97M, P97N,P97Q, P97R, P97S, P97T, P97V, P97W, P97Y, S98*, S98A, S98D, S98E, S98F,S98G, S98H, S98I, S98K, S98L, S98M, S98N, S98P, S98Q, S98R, S98T, S98V,S98W, S98Y, W99*, W99A, W99D, W99E, W99F, W99G, W99H, W99I, W99K, W99L,W99M, W99N, W99P, W99Q, W99R, W99S, W99T, W99V, W99Y, S100*, S100A,S100D, S100E, S100F, S100G, S100H, S100I, S100K, S100L, S100M, S100N,S100P, S100Q, S100R, S100T, S100V, S100W, S100Y, A101*, A101D, A101E,A101F, A101G, A101H, A101I, A101K, A101L, A101M, A101N, A101P, A101Q,A101R, A101S, A101T, A101V, A101W, A101Y, Q102*, Q102A, Q102D, Q102E,Q102F, Q102G, Q102H, Q102I, Q102K, Q102L, Q102M, Q102N, Q102P, Q102R,Q102S, Q102T, Q102V, Q102W, Q102Y, D103*, D103A, D103E, D103F, D103G,D103H, D103I, D103K, D103L, D103M, D103N, D103P, D103Q, D103R, D103S,D103T, D103V, D103W, D103Y, F104*, F104A, F104D, F104E, F104G, F104H,F104I, F104K, F104L, F104M, F104N, F104P, F104Q, F104R, F104S, F104T,F104V, F104W, F104Y, K105*, K105A, K105D, K105E, K105F, K105G, K105H,K105I, K105L, K105M, K105N, K105P, K105Q, K105R, K105S, K105T, K105V,K105W, K105Y, S106*, S106A, S106D, S106E, S106F, S106G, S106H, S106I,S106K, S106L, S106M, S106N, S106P, S106Q, S106R, S106T, S106V, S106W,S106Y, P107*, P107A, P107D, P107E, P107F, P107G, P107H, P107I, P107K,P107L, P107M, P107N, P107Q, P107R, P107S, P107T, P107V, P107W, P107Y,E108*, E108A, E108D, E108F, E108G, E108H, E108I, E108K, E108L, E108M,E108N, E108P, E108Q, E108R, E108S, E108T, E108V, E108W, E108Y, E109*,E109A, E109D, E109F, E109G, E109H, E109I, E109K, E109L, E109M, E109N,E109P, E109Q, E109R, E109S, E109T, E109V, E109W, E109Y, Y110*, Y110A,Y110D, Y110E, Y110F, Y110G, Y110H, Y110I, Y110K, Y110L, Y110M, Y110N,Y110P, Y110Q, Y110R, Y110S, Y110T, Y110V, Y110W, A111*, A111D, A111E,A111F, A111G, A111H, A111I, A111K, A111L, A111M, A111N, A111P, A111Q,A111R, A111S, A111T, A111V, A111W, A111Y, F112*, F112A, F112D, F112E,F112G, F112H, F112I, F112K, F112L, F112M, F112N, F112P, F112Q, F112R,F112S, F112T, F112V, F112W, F112Y, A113*, A113D, A113E, A113F, A113G,A113H, A113I, A113K, A113L, A113M, A113N, A113P, A113Q, A113R, A113S,A113T, A113V, A113W, A113Y, S114*, S114A, S114D, S114E, S114F, S114G,S114H, S114I, S114K, S114L, S114M, S114N, S114P, S114Q, S114R, S114T,S114V, S114W, S114Y, S115*, S115A, S115D, S115E, S115F, S115G, S115H,S115I, S115K, S115L, S115M, S115N, S115P, S115Q, S115R, S115T, S115V,S115W, S115Y, L116*, L116A, L116D, L116E, L116F, L116G, L116H, L116I,L116K, L116M, L116N, L116P, L116Q, L116R, L116S, L116T, L116V, L116W,L116Y, Q117*, Q117A, Q117D, Q117E, Q117F, Q117G, Q117H, Q117I, Q117K,Q117L, Q117M, Q117N, Q117P, Q117R, Q117S, Q117T, Q117V, Q117W, Q117Y,G118*, G118A, G118D, G118E, G118F, G118H, G118I, G118K, G118L, G118M,G118N, G118P, G118Q, G118R, G118S, G118T, G118V, G118W, G118Y, G119*,G119A, G119D, G119E, G119F, G119H, G119I, G119K, G119L, G119M, G119N,G119P, G119Q, G119R, G119S, G119T, G119V, G119W, G119Y, T120*, T120A,T120D, T120E, T120F, T120G, T120H, T120I, T120K, T120L, T120M, T120N,T120P, T120Q, T120R, T120S, T120V, T120W, T120Y, N121*, N121A, N121D,N121E, N121F, N121G, N121H, N121I, N121K, N121L, N121M, N121P, N121Q,N121R, N121S, N121T, N121V, N121W, N121Y, A122*, A122D, A122E, A122F,A122G, A122H, A122I, A122K, A122L, A122M, A122N, A122P, A122Q, A122R,A122S, A122T, A122V, A122W, A122Y, I123*, I123A, I123D, I123E, I123F,I123G, I123H, I123K, I123L, I123M, I123N, I123P, I123Q, I123R, I123S,I123T, I123V, I123W, I123Y, L124*, L124A, L124D, L124E, L124F, L124G,L124H, L124I, L124K, L124M, L124N, L124P, L124Q, L124R, L124S, L124T,L124V, L124W, L124Y, A125*, A125D, A125E, A125F, A125G, A125H, A125I,A125K, A125L, A125M, A125N, A125P, A125Q, A125R, A125S, A125T, A125V,A125W, A125Y, P126*, P126A, P126D, P126E, P126F, P126G, P126H, P126I,P126K, P126L, P126M, P126N, P126Q, P126R, P126S, P126T, P126V, P126W,P126Y, V127*, V127A, V127D, V127E, V127F, V127G, V127H, V127I, V127K,V127L, V127M, V127N, V127P, V127Q, V127R, V127S, V127T, V127W, V127Y,N128*, N128A, N128D, N128E, N128F, N128G, N128H, N128I, N128K, N128L,N128M, N128P, N128Q, N128R, N128S, N128T, N128V, N128W, N128Y, L129*,L129A, L129D, L129E, L129F, L129G, L129H, L129I, L129K, L129M, L129N,L129P, L129Q, L129R, L129S, L129T, L129V, L129W, L129Y, A130*, A130D,A130E, A130F, A130G, A130H, A130I, A130K, A130L, A130M, A130N, A130P,A130Q, A130R, A130S, A130T, A130V, A130W, A130Y, S131*, S131A, S131D,S131E, S131F, S131G, S131H, S131I, S131K, S131L, S131M, S131N, S131P,S131Q, S131R, S131T, S131V, S131W, S131Y, Q132*, Q132A, Q132D, Q132E,Q132F, Q132G, Q132H, Q132I, Q132K, Q132L, Q132M, Q132N, Q132P, Q132R,Q132S, Q132T, Q132V, Q132W, Q132Y, N133*, N133A, N133D, N133E, N133F,N133G, N133H, N133I, N133K, N133L, N133M, N133P, N133Q, N133R, N133S,N133T, N133V, N133W, N133Y, S134*, S134A, S134D, S134E, S134F, S134G,S134H, S134I, S134K, S134L, S134M, S134N, S134P, S134Q, S134R, S134T,S134V, S134W, S134Y, Q135*, Q135A, Q135D, Q135E, Q135F, Q135G, Q135H,Q135I, Q135K, Q135L, Q135M, Q135N, Q135P, Q135R, Q135S, Q135T, Q135V,Q135W, Q135Y, G136*, G136A, G136D, G136E, G136F, G136H, G136I, G136K,G136L, G136M, G136N, G136P, 3p G136Q, G136R, G136S, G136T, G136V, G136W,G136Y, G137*, G137A, G137D, G137E, G137F, G137H, G137I, G137K, G137L,G137M, G137N, G137P, G137Q, G137R, G137S, G137T, G137V, G137W, G137Y,V138*, V138A, V138D, V138E, V138F, V138G, V138H, V138I, V138K, V138L,V138M, V138N, V138P, V138Q, V138R, V138S, V138T, V138W, V138Y, L139*,L139A, L139D, L139E, L139F, L139G, L139H, L139I, L139K, L139M, L139N,L139P, L139Q, L139R, L139S, L139T, L139V, L139W, L139Y, N140*, N140A,N140D, N140E, N140F, N140G, N140H, N140I, N140K, N140L, N140M, N140P,N140Q, N140R, N140S, N140T, N140V, N140W, N140Y, G141*, G141A, G141D,G141E, G141F, G141H, G141I, G141K, G141L, G141M, G141N, G141P, G141Q,G141R, G141S, G141T, G141V, G141W, G141Y, F142*, F142A, F142D, F142E,F142G, F142H, F142I, F142K, F142L, F142M, F142N, F142P, F142Q, F142R,F142S, F142T, F142V, F142W, F142Y, Y143*, Y143A, Y143D, Y143E, Y143F,Y143G, Y143H, Y143I, Y143K, Y143L, Y143M, Y143N, Y143P, Y143Q, Y143R,Y143S, Y143T, Y143V, Y143W, S144*, S144A, S144D, S144E, S144F, S144G,S144H, S144I, S144K, S144L, S144M, S144N, S144P, S144Q, S144R, S144T,S144V, S144W, S144Y, A145*, A145D, A145E, A145F, A145G, A145H, A145I,A145K, A145L, A145M, A145N, A145P, A145Q, A145R, A145S, A145T, A145V,A145W, A145Y, N146*, N146A, N146D, N146E, N146F, N146G, N146H, N146I,N146K, N146L, N146M, N146P, N146Q, N146R, N146S, N146T, N146V, N146W,N146Y, K147*, K147A, K147D, K147E, K147F, K147G, K147H, K147I, K147L,K147M, K147N, K147P, K147Q, K147R, K147S, K147T, K147V, K147W, K147Y,V148*, V148A, V148D, V148E, V148F, V148G, V148H, V148I, V148K, V148L,V148M, V148N, V148P, V148Q, V148R, V148S, V148T, V148W, V148Y, A149*,A149D, A149E, A149F, A149G, A149H, A149I, A149K, A149L, A149M, A149N,A149P, A149Q, A149R, A149S, A149T, A149V, A149W, A149Y, Q150*, Q150A,Q150D, Q150E, Q150F, Q150G, Q150H, Q150I, Q150K, Q150L, Q150M, Q150N,Q150P, Q150R, Q150S, Q150T, Q150V, Q150W, Q150Y, F151*, F151A, F151D,F151E, F151G, F151H, F151I, F151K, F151L, F151M, F151N, F151P, F151Q,F151R, F151S, F151T, F151V, F151W, F151Y, D152*, D152A, D152E, D152F,D152G, D152H, D152I, D152K, D152L, D152M, D152N, D152P, D152Q, D152R,D152S, D152T, D152V, D152W, D152Y, P153*, P153A, P153D, P153E, P153F,P153G, P153H, P153I, P153K, P153L, 26P153M, P153N, P153Q, P153R, P153S,P153T, P153V, P153W, P153Y, S154*, S154A, S154D, S154E, S154F, S154G,S154H, S154I, S154K, S154L, S154M, S154N, S154P, S154Q, S154R, S154T,S154V, S154W, S154Y, K155*, K155A, K155D, K155E, K155F, K155G, K155H,K155I, K155L, K155M, K155N, K155P, K155Q, K155R, K155S, K155T, K155V,K155W, K155Y, P156*, P156A, P156D, P156E, P156F, P156G, P156H, P156I,P156K, P156L, P156M, P156N, P156Q, P156R, 3p P156S, P156T, P156V, P156W,P156Y, Q157*, Q157A, Q157D, Q157E, Q157F, Q157G, Q157H, Q157I, Q157K,Q157L, Q157M, Q157N, Q157P, Q157R, Q157S, Q157T, Q157V, Q157W, Q157Y,Q158*, Q158A, Q158D, Q158E, Q158F, Q158G, Q158H, Q158I, Q158K, Q158L,Q158M, Q158N, Q158P, Q158R, Q158S, Q158T, Q158V, Q158W, Q158Y, T159*,T159A, T159D, T159E, T159F, T159G, T159H, T159I, T159K, T159L, T159M,T159N, T159P, T159Q, T159R, T159S, T159V, T159W, T159Y, K160*, K160A,K160D, K160E, K160F, K160G, K160H, K160I, K160L, K160M, K160N, K160P,K160Q, K160R, K160S, K160T, K160V, K160W, K160Y, G161*, G161A, G161D,G161E, G161F, G161H, G161I, G161K, G161L, G161M, G161N, G161P, G161Q,G161R, G161S, G161T, G161V, G161W, G161Y, T162A, T162D, T162E, T162F,T162G, T162H, T162I, T162K, T162L, T162M, T162N, T162P, T162Q, T162R,T162S, T162T, T162V, T162W, T162Y, W163*, W163A, W163D, W163E, W163F,W163G, W163H, W163I, W163K, W163L, W163M, W163N, W163P, W163Q, W163R,W163S, W163T, W163V, W163Y, F164*, F164A, F164D, F164E, F164G, F164H,F164I, F164K, F164L, F164M, F164N, F164P, F164Q, F164R, F164S, F164T,F164V, F164W, F164Y, Q165*, Q165A, Q165D, Q165E, Q165F, Q165G, Q165H,Q165I, Q165K, Q165L, Q165M, Q165N, Q165P, Q165R, Q165S, Q165T, Q165V,Q165W, Q165Y, I166*, I166A, I166D, I166E, I166F, I166G, I166H, I166K,I166L, I166M, I166N, I166P, I166Q, I166R, I166S, I166T, I166V, I166W,I166Y, T167*, T167A, T167D, T167E, T167F, T167G, T167H, T167I, T167K,T167L, T167M, T167N, T167P, T167Q, T167R, T167S, T167V, T167W, T167Y,K168*, K168A, K168D, K168E, K168F, K168G, K168H, K168I, K168L, K168M,K168N, K168P, K168Q, K168R, K168S, K168T, K168V, K168W, K168Y, F169*,F169A, F169D, F169E, F169G, F169H, F169I, F169K, F169L, F169M, F169N,F169P, F169Q, F169R, F169S, F169T, F169V, F169W, F169Y, T170*, T170A,T170D, T170E, T170F, T170G, T170H, T170I, T170K, T170L, T170M, T170N,T170P, T170Q, T170R, T170S, T170V, T170W, T170Y, G171*, G171A, G171D,G171E, G171F, G171H, G171I, G171K, G171L, G171M, G171N, G171P, G171Q,G171R, G171S, G171T, G171V, G171W, G171Y, A172*, A172D, A172E, A172F,A172G, A172H, A172I, A172K, A172L, A172M, A172N, A172P, A172Q, A172R,A172S, A172T, A172V, A172W, A172Y, A173*, A173D, A173E, A173F, A173G,A173H, A173I, A173K, A173L, A173M, A173N, A173P, A173Q, A173R, A173S,A173T, A173V, A173W, A173Y, G174*, G174A, G174A, G174D, G174DG174A,G174D, G174E, G174E, G174F, G174FG174E, G174F, G174H, G174H, G174I,G1741G174H, G174I, G174K, G174K, G174L, G174LG174K, G174L, G174M, G174M,G174N, G174N, G174M, G174N, G174P, G174P, G174Q, G174QG174P, G174Q,G174R, G174R, G174S, G174SG174R, G174S, G174T, G174T, G174V, G174VG174T,G174V, G174W, G174W, G174Y, G174YG174W, G174Y, P175*, P175A, P175D,P175E, P175F, P175G, P175H, P175I, P175K, P175L, P175M, P175N, P175Q,P175R, P175S, P175T, P175V, P175W, P175Y, Y176*, Y176A, Y176D, Y176E,Y176F, Y176G, Y176H, Y176I, Y176K, Y176L, Y176M, Y176N, Y176P, Y176Q,Y176R, Y176S, Y176T, Y176V, Y176W, K178*, K178A, K178D, K178E, K178F,K178G, K178H, K178I, K178L, K178M, K178N, K178P, K178Q, K178R, K178S,K178T, K178V, K178W, K178Y, A179*, A179D, A179E, A179F, A179G, A179H,A179I, A179K, A179L, A179M, A179N, A179P, A179Q, A179R, A179S, A179T,A179V, A179W, A179Y, L180*, L180A, L180D, L180E, L180F, L180G, L180H,L180I, L180K, L180M, L180N, L180P, L180Q, L180R, L180S, L180T, L180V,L180W, L180Y, G181*, G181A, G181D, G181E, G181F, G181H, G181I, G181K,G181L, G181M, G181N, G181P, G181Q, G181R, G181S, G181T, G181V, G181W,G181Y, S182*, S182A, S182D, S182E, S182F, S182G, S182H, S182I, S182K,S182L, S182M, S182N, S182P, S182Q, S182R, S182T, S182V, S182W, S182Y,N183*, N183A, N183D, N183E, N183F, N183G, N183H, N183I, N183K, N183L,N183M, N183P, N183Q, N183R, N183S, N183T, N183V, N183W, N183Y, D184*,D184A, D184E, D184F, D184G, D184H, D184I, D184K, D184L, D184M, D184N,D184P, D184Q, D184R, D184S, D184T, D184V, D184W, D184Y, K185*, K185A,K185D, K185E, K185F, K185G, K185H, K185I, K185L, K185M, K185N, K185P,K185Q, K185R, K185S, K185T, K185V, K185W, K185Y, S186*, S186A, S186D,S186E, S186F, S186G, S186H, S186I, S186K, S186L, S186M, S186N, S186P,S186Q, S186R, S186T, S186V, S186W, S186Y, V187*, V187A, V187D, V187E,V187F, V187G, V187H, V187I, V187K, V187L, V187M, V187N, V187P, V187Q,V187R, V187S, V187T, V187W, V187Y, D189*, D189A, D189E, D189F, D189G,D189H, D189I, D189K, D189L, D189M, D189N, D189P, D189Q, D189R, D189S,D189T, D189V, D189W, D189Y, K190*, K190A, K190D, K190E, K190F, K190G,K190H, K190I, K190L, K190M, K190N, K190P, K190Q, K190R, K190S, K190T,K190V, K190W, K190Y, N191*, N191A, N191D, N191E, N191F, N191G, N191H,N191I, N191K, N191L, N191M, N191P, N191Q, N191R, N191S, N191T, N191V,N191W, N191Y, K192*, K192A, K192D, K192E, K192F, K192G, K192H, K192I,K192L, K192M, K192N, K192P, K192Q, K192R, K192S, K192T, K192V, K192W,K192Y, N193*, N193A, N193D, N193E, N193F, N193G, N193H, N193I, N193K,N193L, N193M, N193P, N193Q, N193R, N193S, N193T, N193V, N193W, N193Y,I194*, I194A, I194D, I194E, I194F, I194G, I194H, I194K, I194L, I194M,I194N, I194P, I194Q, I194R, I194S, I194T, I194V, I194W, I194Y, A195*,A195D, A195E, A195F, A195G, A195H, A195I, A195K, A195L, A195M, A195N,A195P, A195Q, A195R, A195S, A195T, A195V, A195W, A195Y, G196*, G196A,G196D, G196E, G196F, G196H, G196I, G196K, G196L, G196M, G196N, G196P,G196Q, G196R, G196S, G196T, G196V, G196W, G196Y, D197*, D197A, D197E,D197F, D197G, D197H, D197I, D197K, D197L, D197M, D197N, D197P, D197Q,D197R, D197S, D197T, D197V, D197W, D197Y, W198*, W198A, W198D, W198E,W198F, W198G, W198H, W198I, W198K, W198L, W198M, W198N, W198P, W198Q,W198R, W198S, W198T, W198V, W198Y, G199*, G199A, G199D, G199E, G199F,G199H, G199I, G199K, G199L, G199M, G199N, G199P, G199Q, G199R, G199S,G199T, G199V, G199W, G199Y, F200*, F200A, F200D, F200E, F200G, F200H,F200I, F200K, F200L, F200M, F200N, F200P, F200Q, F200R, F200S, F200T,F200V, F200W, F200Y, D201*, D201A, D201E, D201F, D201G, D201H, D201I,D201K, D201L, D201M, D201N, D201P, D201Q, D201R, D201S, D201T, D201V,D201W, D201Y, P202*, P202A, P202D, P202E, P202F, P202G, P202H, P202I,P202K, P202L, P202M, P202N, P202Q, P202R, P202S, P202T, P202V, P202W,P202Y, A203*, A203D, A203E, A203F, A203G, A203H, A203I, A203K, A203L,A203M, A203N, A203P, A203Q, A203R, A203S, A203T, A203V, A203W, A203Y,K204*, K204A, K204D, K204E, K204F, K204G, K204H, K204I, K204L, K204M,K204N, K204P, K204Q, K204R, K204S, K204T, K204V, K204W, K204Y, W205*,W205A, W205D, W205E, W205F, W205G, W205H, W205I, W205K, W205L, W205M,W205N, W205P, W205Q, W205R, W205S, W205T, W205V, W205Y, A206*, A206D,A206E, A206F, A206G, A206H, A206I, A206K, A206L, A206M, A206N, A206P,A206Q, A206R, A206S, A206T, A206V, A206W, A206Y, Y207*, Y207A, Y207D,Y207E, Y207F, Y207G, Y207H, Y207I, Y207K, Y207L, Y207M, Y207N, Y207P,Y207Q, Y207R, Y207S, Y207T, Y207V, Y207W, Q208*, Q208A, Q208D, Q208E,Q208F, Q208G, Q208H, Q208I, Q208K, Q208L, Q208M, Q208N, Q208P, Q208R,Q208S, Q208T, Q208V, Q208W, Q208Y, Y209*, Y209A, Y209D, Y209E, Y209F,Y209G, Y209H, Y209I, Y209K, Y209L, Y209M, Y209N, Y209P, Y209Q, Y209R,Y209S, Y209T, Y209V, Y209W, D210*, D210A, D210E, D210F, D210G, D210H,D210I, D210K, D210L, D210M, D210N, D210P, D210Q, D210R, D210S, D210T,D210V, D210W, D210Y, E211*, E211A, E211D, E211F, E211G, E211H, E211I,E211K, E211L, E211M, E211N, E211P, E211Q, E211R, E211S, E211T, E211V,E211W, E211Y, K212*, K212A, K212D, K212E, K212F, K212G, K212H, K212I,K212L, K212M, K212N, K212P, K212Q, K212R, K212S, K212T, K212V, K212W,K212Y, N213*, N213A, N213D, N213E, N213F, N213G, N213H, N213I, N213K,N213L, N213M, N213P, N213Q, N213R, N213S, N213T, N213V, N213W, N213Y,N214*, N214A, N214D, N214E, N214F, N214G, N214H, N214I, N214K, N214L,N214M, N214P, N214Q, N214R, N214S, N214T, N214V, N214W, N214Y, K215*,K215A, K215D, K215E, K215F, K215G, K215H, K215I, K215L, K215M, K215N,K215P, K215Q, K215R, K215S, K215T, K215V, K215W, K215Y, F216*, F216A,F216D, F216E, F216G, F216H, F216I, F216K, F216L, F216M, F216N, F216P,F216Q, F216R, F216S, F216T, F216V, F216W, F216Y, N217*, N217A, N217D,N217E, N217F, N217G, N217H, N217I, N217K, N217L, N217M, N217P, N217Q,N217R, N217S, N217T, N217V, N217W, N217Y, Y218*, Y218A, Y218D, Y218E,Y218F, Y218G, Y218H, Y218I, Y218K, Y218L, Y218M, Y218N, Y218P, Y218Q,Y218R, Y218S, Y218T, Y218V, Y218W, V219*, V219A, V219D, V219E, V219F,V219G, V219H, V219I, V219K, V219L, V219M, V219N, V219P, V219Q, V219R,V219S, V219T, V219W, V219Y, G220*, G220A, G220D, G220E, G220F, G220H,G220I, G220K, G220L, G220M, G220N, G220P, G220Q, G220R, G220S, G220T,G220V, G220W, G220Y, K221*, K221A, K221D, K221E, K221F, K221G, K221H,K221I, K221L, K221M, K221N, K221P, K221Q, K221R, K221S, K221T, K221V,K221W and K221Y, wherein the variant has a sequence identity to SEQ IDNO: 1 of at least 60% and the variant has DNase activity.3. The variant of any of embodiments 1-2, which has an improveddetergent stability compared to the parent or compared to the DNase withSEQ ID NO: 1.4. The variant of any of embodiments 1-3, wherein the variant has atleast 80%, at least 85%, at least 90%, at least 95%, at least 96%, atleast 97%, at least 98%, at least 99 sequence identity to the maturepolypeptide of SEQ ID NO: 1.5. The variant of any of embodiments 1-4, wherein the total number ofalterations compared to SEQ ID NO: 1 is 1-20, e.g. 1-10 and 1-5, such as1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 alterations.6. A detergent composition comprising a variant according to any ofembodiments 1 to 5.7. The detergent composition of embodiment 6, further comprising one ormore detergent components.8. The detergent composition according to any of embodiments 6-7,further comprising one or more additional enzymes selected from thegroup comprising of proteases, amylases, lipases, cutinases, cellulases,endoglucanases, xyloglucanases, pectinases, pectin lyases, xanthanases,peroxidaes, haloperoxygenases, catalases and mannanases, or any mixturethereof.9. The detergent composition according to any of embodiments 6-8 in formof a bar, a homogenous tablet, a tablet having two or more layers, apouch having one or more compartments, a regular or compact powder, agranule, a paste, a gel, or a regular, compact or concentrated liquid.10. Use of a detergent composition according to any of embodiments 6-9in a cleaning process, such as laundry or hard surface cleaning such asdish wash.11. A method for obtaining a DNase variant, comprising introducing intoa parent DNase an alteration at one or more positions corresponding topositions 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18,19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36,37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 53, 54, 55,56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73,74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91,92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107,108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121,122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135,136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149,150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163,164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 174, 175, 176, 177,178, 179, 180, 181, 182, 183, 185, 186, 187, 189, 190, 191, 192, 193,194, 195, 196, 197, 198, 199, 200, 201, 202, 203, 204, 205, 206, 207,208, 209, 210, 211, 212, 213, 214, 215, 216, 217, 218, 219, 220 or 221of SEQ ID NO 1, wherein the variant has an amino acid sequence which isat least 60% identical to SEQ ID NO 1, and recovering the variant.12. The method of embodiment 11, wherein the variant comprises two,three, four or five alterations compared to SEQ ID NO: 1.13. The method according to any of embodiments 11 or 12, wherein thevariant comprises one or more alterations selected from the groupconsisting of V1*, V1A, V1D, V1E, V1F, V1G, V1H, V1I, V1K, V1L, V1M,V1N, V1P, V1Q, V1R, V1S, V1T, V1W, V1Y, P2*, P2A, P2D, P2E, P2F, P2G,P2H, P2I, P2K, P2L, P2M, P2N, P2Q, P2R, P2S, P2T, P2V, P2W, P2Y, V3*,V3A, V3C, V3D, V3E, V3F, V3G, V3H, V3I, V3K, V3L, V3M, V3N, V3P, V3R,V3S, V3T, V3W, V3Y, N4*, N4A, N4D, N4E, N4F, N4G, N4H, N4I, N4K, N4L,N4M, N4P, N4Q, N4R, N4S, N4T, N4V, N4W, N4Y, P5*, P5A, P5D, P5E, P5F,P5G, P5H, P5I, P5K, P5L, P5M, P5N, P5Q, P5R, P5S, P5T, P5V, P5W, P5Y,E6*, E6A, E6D, E6F, E6G, E6H, E6I, E6K, E6L, E6M, E6N, E6P, E6Q, E6R,E6S, E6T, E6V, E6W, E6Y, P7*, P7A, P7D, P7E, P7F, P7G, P7H, P7I, P7K,P7L, P7M, P7N, P7Q, P7R, P7S, P7T, P7V, P7W, P7Y, D8*, D8A, D8E, D8F,D8G, D8H, D8I, D8K, D8L, D8M, D8N, D8P, D8Q, D8R, D8S, D8T, D8V, D8W,D8Y, A9*, A9D, A9E, A9F, A9G, A9H, A9I, A9K, A9L, A9M, A9N, A9P, A9Q,A9R, A9S, A9T, A9V, A9W, A9Y, T10*, T10A, T10D, T10E, T10F, T10G, T10H,T10I, T10K, T10L, T10M, T10N, T10P, T10Q, T10R, T10S, T10V, T10W, T10Y,S11*, S11A, S11D, S11E, S11F, S11G, S11H, S11i, S11K, S11L, S11M, S11N,S11P, S11Q, S11R, S11T, S11V, S11W, S11Y, V12*, V12A, V12D, V12E, V12F,V12G, V12H, V12I, V12K, V12L, V12M, V12N, V12P, V12Q, V12R, V12S, V12T,V12W, V12Y, E13*, E13A, E13D, E13F, E13G, E13H, E13I, E13K, E13L, E13M,E13N, E13P, E13Q, E13R, E13S, E13T, E13V, E13W, E13Y, N14*, N14A, N14D,N14E, N14F, N14G, N14H, N14I, N14K, N14L, N14M, N14P, N14Q, N14R, N14S,N14T, N14V, N14W, N14Y, V15*, V15A, V15D, V15E, V15F, V15G, V15H, V15I,V15K, V15L, V15M, V15N, V15P, V15Q, V15R, V15S, V15T, V15W, V15Y, A16*,A16D, A16E, A16F, A16G, A16H, A16I, A16K, A16L, A16M, A16N, A16P, A16Q,A16R, A16S, A16T, A16V, A16W, A16Y, L17*, L17A, L17D, L17E, L17F, L17G,L17H, L17I, L17K, L17M, L17N, L17P, L17Q, L17R, L17S, L17T, L17V, L17W,L17Y, K18*, K18A, K18D, K18E, K18F, K18G, K18H, K18I, K18L, K18M, K18N,K18P, K18Q, K18R, K18S, K18T, K18V, K18W, K18Y, T19*, T19A, T19D, T19E,T19F, T19G, T19H, T19I, T19K, T19L, T19M, T19N, T19P, T19Q, T19R, T19S,T19V, T19W, T19Y, G20*, G20A, G20D, G20E, G20F, G20H, G20I, G20K, G20L,G20M, G20N, G20P, G20Q, G20R, G20S, G20T, G20V, G20W, G20Y, S21*, S21A,S21D, S21E, S21F, S21G, S21H, S21I, S21K, S21L, S21M, S21N, S21P, S21Q,S21R, S21T, S21V, S21W, S21Y, G22*, G22A, G22D, G22E, G22F, G22H, G22I,G22K, G22L, G22M, G22N, G22P, G22Q, G22R, G22S, G22T, G22V, G22W, G22Y,D23*, D23A, D23E, D23F, D23G, D23H, D23I, D23K, D23L, D23M, D23N, D23P,D23Q, D23R, D23S, D23T, D23V, D23W, D23Y, S24*, S24A, S24D, S24E, S24F,S24G, S24H, S24I, S24K, S24L, S24M, S24N, S24P, S24Q, S24R, S24T, S24V,S24W, S24Y, Q25*, Q25A, Q25D, Q25E, Q25F, Q25G, Q25H, Q25I, Q25K, Q25L,Q25M, Q25N, Q25P, Q25R, Q25S, Q25T, Q25V, Q25W, Q25Y, S26*, S26A, S26D,S26E, S26F, S26G, S26H, S26I, S26K, S26L, S26M, S26N, S26P, S26Q, S26R,S26T, S26V, S26W, S26Y, D27*, D27A, D27E, D27F, D27G, D27H, D27I, D27K,D27L, D27M, D27N, D27P, D27Q, D27R, D27S, D27T, D27V, D27W, D27Y, P28*,P28A, P28D, P28E, P28F, P28G, P28H, P28I, P28K, P28L, P28M, P28N, P28Q,P28R, P28S, P28T, P28V, P28W, P28Y, I29*, I29A, I29D, I29E, I29F, I29G,I29H, I29K, I29L, I29M, I29N, I29P, I29Q, I29R, I29S, I29T, I29V, I29W,I29Y, K30*, K30A, K30D, K30E, K30F, K30G, K30H, K30I, K30L, K30M, K30N,K30P, K30Q, K30R, K30S, K30T, K30V, K30W, K30Y, A31*, A31D, A31E, A31F,A31G, A31H, A31I, A31K, A31L, A31M, A31N, A31P, A31Q, A31R, A31S, A31T,A31V, A31W, A31Y, D32*, D32A, D32E, D32F, D32G, D32H, D32I, D32K, D32L,D32M, D32N, D32P, D32Q, D32R, D32S, D32T, D32V, D32W, D32Y, L33*, L33A,L33D, L33E, L33F, L33G, L33H, L33I, L33K, L33M, L33N, L33P, L33Q, L33R,L33S, L33T, L33V, L33W, L33Y, E34*, E34A, E34D, E34F, E34G, E34H, E34I,E34K, E34L, E34M, E34N, E34P, E34Q, E34R, E34S, E34T, E34V, E34W, E34Y,V35*, V35A, V35D, V35E, V35F, V35G, V35H, V35I, V35K, V35L, V35M, V35N,V35P, V35Q, V35R, V35S, V35T, V35W, V35Y, K36*, K36A, K36D, K36E, K36F,K36G, K36H, K36I, K36L, K36M, K36N, K36P, K36Q, K36R, K36S, K36T, K36V,K36W, K36Y, G37*, G37A, G37D, G37E, G37F, G37H, G37I, G37K, G37L, G37M,G37N, G37P, G37Q, G37R, G37S, G37T, G37V, G37W, G37Y, Q38*, Q38A, Q38D,Q38E, Q38F, Q38G, Q38H, Q38I, Q38K, Q38L, Q38M, Q38N, Q38P, Q38R, Q38S,Q38T, Q38V, Q38W, Q38Y, S39*, S39A, S39D, S39E, S39F, S39G, S39H, S39I,S39K, S39L, S39M, S39N, S39P, S39Q, S39R, S39T, S39V, S39W, S39Y, A40*,A40D, A40E, A40F, A40G, A40H, A40I, A40K, A40L, A40M, A40N, A40P, A40Q,A40R, A40S, A40T, A40V, A40W, A40Y, L41*, L41A, L41D, L41E, L41F, L41G,L41H, L41I, L41K, L41M, L41N, L41P, L41Q, L41R, L41S, L41T, L41V, L41W,L41Y, P42*, P42A, P42D, P42E, P42F, P42G, P42H, P42I, P42K, P42L, P42M,P42N, P42Q, P42R, P42S, P42T, P42V, P42W, P42Y, F43*, F43A, F43D, F43E,F43G, F43H, F43I, F43K, F43L, F43M, F43N, F43P, F43Q, F43R, F43S, F43T,F43V, F43W, F43Y, D44*, D44A, D44E, D44F, D44G, D44H, D44I, D44K, D44L,D44M, D44N, D44P, D44Q, D44R, D44S, D44T, D44V, D44W, D44Y, V45*, V45A,V45D, V45E, V45F, V45G, V45H, V45I, V45K, V45L, V45M, V45N, V45P, V45Q,V45R, V45S, V45T, V45W, V45Y, D46*, D46A, D46E, D46F, D46G, D46H, D46I,D46K, D46L, D46M, D46N, D46P, D46Q, D46R, D46S, D46T, D46V, D46W, D46Y,C47A, C47D, C47E, C47F, C47G, C47H, C47I, C47K, C47L, C47M, C47N, C47P,C47Q, C47R, C47S, C47T, C47V, C47W, C47Y, W48*, W48A, W48D, W48E, W48F,W48G, W48H, W48I, W48K, W48L, W48M, W48N, W48P, W48Q, W48R, W48S, W48T,W48V, W48Y, A49*, A49D, A49E, A49F, A49G, A49H, A49I, A49K, A49L, A49M,A49N, A49P, A49Q, A49R, A49S, A49T, A49V, A49W, A49Y, I50*, I50A, I50D,150E, I50F, I50G, I50H, I50K, I50L, I50M, I50N, I50P, I50Q, I50R, I50S,I50T, I50V, I50W, I50Y, L51*, L51A, L51D, L51E, L51F, L51G, L51H, L51I,L51K, L51M, L51N, L51P, L51Q, L51R, L51S, L51T, L51V, L51W, L51Y, K53*,K53A, K53D, K53E, K53F, K53G, K53H, K53I, K53L, K53M, K53N, K53P, K53Q,K53R, K53S, K53T, K53V, K53W, K53Y, G54*, G54A, G54D, G54E, G54F, G54H,G54I, G54K, G54L, G54M, G54N, G54P, G54Q, G54R, G54S, G54T, G54V, G54W,G54Y, A55*, A55D, A55E, A55F, A55G, A55H, A55I, A55K, A55L, A55M, A55N,A55P, A55Q, A55R, A55S, A55T, A55V, A55W, A55Y, P56*, P56A, P56D, P56E,P56F, P56G, P56H, P56I, P56K, P56L, P56M, P56N, P56Q, P56R, P56S, P56T,P56V, P56W, P56Y, N57*, N57A, N57D, N57E, N57F, N57G, N57H, N57I, N57K,N57L, N57M, N57P, N57Q, N57R, N57S, N57T, N57V, N57W, N57Y, V58*, V58A,V58D, V58E, V58F, V58G, V58H, V58I, V58K, V58L, V58M, V58N, V58P, V58Q,V58R, V58S, V58T, V58W, V58YL59*, L59A, L59A, L59D, L59D, L59A, L59D,L59E, L59E, L59F, L59F, L59E, L59F, L59G, L59G, L59H, L59H, L59G, L59H,L59I, L59I, L59K, L59K, L59I, L59K, L59M, L59ML59M, L59N, L59N, L59N,L59P, L59P, L59Q, L59Q, L59P, L59Q, L59R, L59R, L59S, L59S, L59R, L59S,L59T, L59T, L59V, L59V, L59T, L59V, L59W, L59W, L59Y, L59YL59W, L59Y,Q60*, Q60A, Q60D, Q60E, Q60F, Q60G, Q60H, Q60I, Q60K, Q60L, Q60M, Q60N,Q60P, Q60R, Q60S, Q60T, Q60V, Q60W, Q60Y, R61*, R61A, R61D, R61E, R61F,R61G, R61H, R61I, R61K, R61L, R61M, R61N, R61P, R61Q, R61S, R61T, R61V,R61W, R61Y, V62*, V62A, V62D, V62E, V62F, V62G, V62H, V62I, V62K, V62L,V62M, V62N, V62P, V62Q, V62R, V62S, V62T, V62W, V62Y, N63*, N63A, N63D,N63E, N63F, N63G, N63H, N63I, N63K, N63L, N63M, N63P, N63Q, N63R, N63S,N63T, N63V, N63W, N63Y, E64*, E64A, E64D, E64F, E64G, E64H, E64I, E64K,E64L, E64M, E64N, E64P, E64Q, E64R, E64S, E64T, E64V, E64W, E64Y, K65*,K65A, K65D, K65E, K65F, K65G, K65H, K65I, K65L, K65M, K65N, K65P, K65Q,K65R, K65S, K65T, K65V, K65W, K65Y, T66*, T66A, T66D, T66E, T66F, T66G,T66H, T66I, T66K, T66L, T66M, T66N, T66P, T66Q, T66R, T66S, T66V, T66W,T66Y, K67*, K67A, K67D, K67E, K67F, K67G, K67H, K67I, K67L, K67M, K67N,K67P, K67Q, K67R, K67S, K67T, K67V, K67W, K67Y, N68*, N68A, N68D, N68E,N68F, N68G, N68H, N68I, N68K, N68L, N68M, N68P, N68Q, N68R, N68S, N68T,N68V, N68W, N68Y, S69*, S69A, S69D, S69E, S69F, S69G, S69H, S69I, S69K,S69L, S69M, S69N, S69P, S69Q, S69R, S69T, S69V, S69W, S69Y, N70*, N70A,N70D, N70E, N70F, N70G, N70H, N70I, N70K, N70L, N70M, N70P, N70Q, N70R,N70S, N70T, N70V, N70W, N70Y, R71*, R71A, R71D, R71E, R71F, R71G, R71H,R71I, R71K, R71L, R71M, R71N, R71P, R71Q, R71S, R71T, R71V, R71W, R71Y,D72*, D72A, D72E, D72F, D72G, D72H, D72I, D72K, D72L, D72M, D72N, D72P,D72Q, D72R, D72S, D72T, D72V, D72W, D72Y, R73*, R73A, R73D, R73E, R73F,R73G, R73H, R73I, R73K, R73L, R73M, R73N, R73P, R73Q, R73S, R73T, R73V,R73W, R73Y, S74*, S74A, S74D, S74E, S74F, S74G, S74H, S74I, S74K, S74L,S74M, S74N, S74P, S74Q, S74R, S74T, S74V, S74W, S74Y, G75*, G75A, G75D,G75E, G75F, G75H, G75I, G75K, G75L, G75M, G75N, G75P, G75Q, G75R, G75S,G75T, G75V, G75W, G75Y, A76*, A76D, A76E, A76F, A76G, A76H, A76I, A76K,A76L, A76M, A76N, A76P, A76Q, A76R, A76S, A76T, A76V, A76W, A76Y, N77*,N77A, N77D, N77E, N77F, N77G, N77H, N77I, N77K, N77L, N77M, N77P, N77Q,N77R, N77S, N77T, N77V, N77W, N77Y, K78*, K78A, K78D, K78E, K78F, K78G,K78H, K78I, K78L, K78M, K78N, K78P, K78Q, K78R, K78S, K78T, K78V, K78W,K78Y, G79*, G79A, G79D, G79E, G79F, G79H, G79I, G79K, G79L, G79M, G79N,G79P, G79Q, G79R, G79S, G79T, G79V, G79W, G79Y, P80*, P80A, P80D, P80E,P80F, P80G, P80H, P80I, P80K, P80L, P80M, P80N, P80Q, P80R, P80S, P80T,P80V, P80W, P80Y, F81*, F81A, F81D, F81E, F81G, F81H, F81I, F81K, F81L,F81M, F81N, F81P, F81Q, F81R, F81S, F81T, F81V, F81W, F81Y, K82*, K82A,K82D, K82E, K82F, K82G, K82H, K82I, K82L, K82M, K82N, K82P, K82Q, K82R,K82S, K82T, K82V, K82W, K82Y, D83*, D83A, D83E, D83F, D83G, D83H, D83I,D83K, D83L, D83M, D83N, D83P, D83Q, D83R, D83S, D83T, D83V, D83W, D83Y,P84*, P84A, P84D, P84E, P84F, P84G, P84H, P84I, P84K, P84L, P84M, P84N,P84Q, P84R, P84S, P84T, P84V, P84W, P84Y, Q85*, Q85A, Q85D, Q85E, Q85F,Q85G, Q85H, Q85I, Q85K, Q85L, Q85M, Q85N, Q85P, Q85R, Q85S, Q85T, Q85V,Q85W, Q85Y, K86*, K86A, K86D, K86E, K86F, K86G, K86H, K86I, K86L, K86M,K86N, K86P, K86Q, K86R, K86S, K86T, K86V, K86W, K86Y, W87*, W87A, W87D,W87E, W87F, W87G, W87H, W87I, W87K, W87L, W87M, W87N, W87P, W87Q, W87R,W87S, W87T, W87V, W87Y, G88*, G88A, G88D, G88E, G88F, G88H, G88I, G88K,G88L, G88M, G88N, G88P, G88Q, G88R, G88S, G88T, G88V, G88W, G88Y, I89*,I89A, I89D, I89E, I89F, I89G, I89H, I89K, I89L, I89M, I89N, I89P, I89Q,I89R, I89S, I89T, I89V, I89W, I89Y, K90*, K90A, K90D, K90E, K90F, K90G,K90H, K90I, K90L, K90M, K90N, K90P, K90Q, K90R, K90S, K90T, K90V, K90W,K90Y, A91*, A91D, A91E, A91F, A91G, A91H, A91I, A91K, A91L, A91M, A91N,A91P, A91Q, A91R, A91S, A91T, A91V, A91W, A91Y, L92*, L92A, L92D, L92E,L92F, L92G, L92H, L92I, L92K, L92M, L92N, L92P, L92Q, L92R, L92S, L92T,L92V, L92W, L92Y, P93*, P93A, P93D, P93E, P93F, P93G, P93H, P93I, P93K,P93L, P93M, P93N, P93Q, P93R, P93S, P93T, P93V, P93W, P93Y, P94*, P94A,P94D, P94E, P94F, P94G, P94H, P94I, P94K, P94L, P94M, P94N, P94Q, P94R,P94S, P94T, P94V, P94W, P94Y, K95*, K95A, K95D, K95E, K95F, K95G, K95H,K95I, K95L, K95M, K95N, K95P, K95Q, K95R, K95S, K95T, K95V, K95W, K95Y,N96*, N96A, N96D, N96E, N96F, N96G, N96H, N96I, N96K, N96L, N96M, N96P,N96Q, N96R, N96S, N96T, N96V, N96W, N96Y, P97*, P97A, P97D, P97E, P97F,P97G, P97H, P97I, P97K, P97L, P97M, P97N, P97Q, P97R, P97S, P97T, P97V,P97W, P97Y, S98*, S98A, S98D, S98E, S98F, S98G, S98H, S98I, S98K, S98L,S98M, S98N, S98P, S98Q, S98R, S98T, S98V, S98W, S98Y, W99*, W99A, W99D,W99E, W99F, W99G, W99H, W99I, W99K, W99L, W99M, W99N, W99P, W99Q, W99R,W99S, W99T, W99V, W99Y, S100*, S100A, S100D, S100E, S100F, S100G, S100H,S100I, S100K, S100L, S100M, S100N, S100P, S100Q, S100R, S100T, S100V,S100W, S100Y, A101*, A101D, A101E, A101F, A101G, A101H, A101I, A101K,A101L, A101M, A101N, A101P, A101Q, A101R, A101S, A101T, A101V, A101W,A101Y, Q102*, Q102A, Q102D, Q102E, Q102F, Q102G, Q102H, Q102I, Q102K,Q102L, Q102M, Q102N, Q102P, Q102R, Q102S, Q102T, Q102V, Q102W, Q102Y,D103*, D103A, D103E, D103F, D103G, D103H, D103I, D103K, D103L, D103M,D103N, D103P, D103Q, D103R, D103S, D103T, D103V, D103W, D103Y, F104*,F104A, F104D, F104E, F104G, F104H, F104I, F104K, F104L, F104M, F104N,F104P, F104Q, F104R, F104S, F104T, F104V, F104W, F104Y, K105*, K105A,K105D, K105E, K105F, K105G, K105H, K105I, K105L, K105M, K105N, K105P,K105Q, K105R, K105S, K105T, K105V, K105W, K105Y, S106*, S106A, S106D,S106E, S106F, S106G, S106H, S106I, S106K, S106L, S106M, S106N, S106P,S106Q, S106R, S106T, S106V, S106W, S106Y, P107*, P107A, P107D, P107E,P107F, P107G, P107H, P107I, P107K, P107L, P107M, P107N, P107Q, P107R,P107S, P107T, P107V, P107W, P107Y, E108*, E108A, E108D, E108F, E108G,E108H, E108I, E108K, E108L, E108M, E108N, E108P, E108Q, E108R, E108S,E108T, E108V, E108W, E108Y, E109*, E109A, E109D, E109F, E109G, E109H,E109I, E109K, E109L, E109M, E109N, E109P, E109Q, E109R, E109S, E109T,E109V, E109W, E109Y, Y110*, Y110A, Y110D, Y110E, Y110F, Y110G, Y110H,Y110I, Y110K, Y110L, Y110M, Y110N, Y110P, Y110Q, Y110R, Y110S, Y110T,Y110V, Y110W, A111*, A111D, A111E, A111F, A111G, A111H, A111I, A111K,A111L, A111M, A111N, A111P, A111Q, A111R, A111S, A111T, A111V, A111W,A111Y, F112*, F112A, F112D, F112E, F112G, F112H, F112I, F112K, F112L,F112M, F112N, F112P, F112Q, F112R, F112S, F112T, F112V, F112W, F112Y,A113*, A113D, A113E, A113F, A113G, A113H, A113I, A113K, A113L, A113M,A113N, A113P, A113Q, A113R, A113S, A113T, A113V, A113W, A113Y, S114*,S114A, S114D, S114E, S114F, S114G, S114H, S114I, S114K, S114L, S114M,S114N, S114P, S114Q, S114R, S114T, S114V, S114W, S114Y, S115*, S115A,S115D, S115E, S115F, S115G, S115H, S115I, S115K, S115L, S115M, S115N,S115P, S115Q, S115R, S115T, S115V, S115W, S115Y, L116*, L116A, L116D,L116E, L116F, L116G, L116H, L116I, L116K, L116M, L116N, L116P, L116Q,L116R, L116S, L116T, L116V, L116W, L116Y, Q117*, Q117A, Q117D, Q117E,Q117F, Q117G, Q117H, Q117I, Q117K, Q117L, Q117M, Q117N, Q117P, Q117R,Q117S, Q117T, Q117V, Q117W, Q117Y, G118*, G118A, G118D, G118E, G118F,G118H, G118I, G118K, G118L, G118M, G118N, G118P, G118Q, G118R, G118S,G118T, G118V, G118W, G118Y, G119*, G119A, G119D, G119E, G119F, G119H,G119I, G119K, G119L, G119M, G119N, G119P, G119Q, G119R, G119S, G119T,G119V, G119W, G119Y, T120*, T120A, T120D, T120E, T120F, T120G, T120H,T120I, T120K, T120L, T120M, T120N, T120P, T120Q, T120R, T120S, T120V,T120W, T120Y, N121*, N121A, N121D, N121E, N121F, N121G, N121H, N121I,N121K, N121L, N121M, N121P, N121Q, N121R, N121S, N121T, N121V, N121W,N121Y, A122*, A122D, A122E, A122F, A122G, A122H, A122I, A122K, A122L,A122M, A122N, A122P, A122Q, A122R, A122S, A122T, A122V, A122W, A122Y,I123*, I123A, I123D, I123E, I123F, I123G, I123H, I123K, I123L, I123M,I123N, I123P, I123Q, I123R, I123S, I123T, I123V, I123W, I123Y, L124*,L124A, L124D, L124E, L124F, L124G, L124H, L124I, L124K, L124M, L124N,L124P, L124Q, L124R, L124S, L124T, L124V, L124W, L124Y, A125*, A125D,A125E, A125F, A125G, A125H, A125I, A125K, A125L, A125M, A125N, A125P,A125Q, A125R, A125S, A125T, A125V, A125W, A125Y, P126*, P126A, P126D,P126E, P126F, P126G, P126H, P126I, P126K, P126L, P126M, P126N, P126Q,P126R, P126S, P126T, P126V, P126W, P126Y, V127*, V127A, V127D, V127E,V127F, V127G, V127H, V127I, V127K, V127L, V127M, V127N, V127P, V127Q,V127R, V127S, V127T, V127W, V127Y, N128*, N128A, N128D, N128E, N128F,N128G, N128H, N128I, N128K, N128L, N128M, N128P, N128Q, N128R, N128S,N128T, N128V, N128W, N128Y, L129*, L129A, L129D, L129E, L129F, L129G,L129H, L129I, L129K, L129M, L129N, L129P, L129Q, L129R, L129S, L129T,L129V, L129W, L129Y, A130*, A130D, A130E, A130F, A130G, A130H, A130I,A130K, A130L, A130M, A130N, A130P, A130Q, A130R, A130S, A130T, A130V,A130W, A130Y, S131*, S131A, S131D, S131E, S131F, S131G, S131H, S131I,S131K, S131L, S131M, S131N, S131P, S131Q, S131R, S131T, S131V, S131W,S131Y, Q132*, Q132A, Q132D, Q132E, Q132F, Q132G, Q132H, Q132I, Q132K,Q132L, Q132M, Q132N, Q132P, Q132R, Q132S, Q132T, Q132V, Q132W, Q132Y,N133*, N133A, N133D, N133E, N133F, N133G, N133H, N133I, N133K, N133L,N133M, N133P, N133Q, N133R, N133S, N133T, N133V, N133W, N133Y, S134*,S134A, S134D, S134E, S134F, S134G, S134H, S134I, S134K, S134L, S134M,S134N, S134P, S134Q, S134R, S134T, S134V, S134W, S134Y, Q135*, Q135A,Q135D, Q135E, Q135F, Q135G, Q135H, Q135I, Q135K, Q135L, Q135M, Q135N,Q135P, Q135R, Q135S, Q135T, Q135V, Q135W, Q135Y, G136*, G136A, G136D,G136E, G136F, G136H, G136I, G136K, G136L, G136M, G136N, G136P, G136Q,G136R, G136S, G136T, G136V, G136W, G136Y, G137*, G137A, G137D, G137E,G137F, G137H, G137I, G137K, G137L, G137M, G137N, G137P, G137Q, G137R,G137S, G137T, G137V, G137W, G137Y, V138*, V138A, V138D, V138E, V138F,V138G, V138H, V138I, V138K, V138L, V138M, V138N, V138P, V138Q, V138R,V138S, V138T, V138W, V138Y, L139*, L139A, L139D, L139E, L139F, L139G,L139H, L139I, L139K, L139M, L139N, L139P, L139Q, L139R, L139S, L139T,L139V, L139W, L139Y, N140*, N140A, N140D, N140E, N140F, N140G, N140H,N140I, N140K, N140L, N140M, N140P, N140Q, N140R, N140S, N140T, N140V,N140W, N140Y, G141*, G141A, G141D, G141E, G141F, G141H, G141I, G141K,G141L, G141M, G141N, G141P, G141Q, G141R, G141S, G141T, G141V, G141W,G141Y, F142*, F142A, F142D, F142E, F142G, F142H, F142I, F142K, F142L,F142M, F142N, F142P, F142Q, F142R, F142S, F142T, F142V, F142W, F142Y,Y143*, Y143A, Y143D, Y143E, Y143F, Y143G, Y143H, Y143I, Y143K, Y143L,Y143M, Y143N, Y143P, Y143Q, Y143R, Y143S, Y143T, Y143V, Y143W, S144*,S144A, S144D, S144E, S144F, S144G, S144H, S144I, S144K, S144L, S144M,S144N, S144P, S144Q, S144R, S144T, S144V, S144W, S144Y, A145*, A145D,A145E, A145F, A145G, A145H, A145I, A145K, A145L, A145M, A145N, A145P,A145Q, A145R, A145S, A145T, A145V, A145W, A145Y, N146*, N146A, N146D,N146E, N146F, N146G, N146H, N146I, N146K, N146L, N146M, N146P, N146Q,N146R, N146S, N146T, N146V, N146W, N146Y, K147*, K147A, K147D, K147E,K147F, K147G, K147H, K147I, K147L, K147M, K147N, K147P, K147Q, K147R,K147S, K147T, K147V, K147W, K147Y, V148*, V148A, V148D, V148E, V148F,V148G, V148H, V148I, V148K, V148L, V148M, V148N, V148P, V148Q, V148R,V148S, V148T, V148W, V148Y, A149*, A149D, A149E, A149F, A149G, A149H,A149I, A149K, A149L, A149M, A149N, A149P, A149Q, A149R, A149S, A149T,A149V, A149W, A149Y, Q150*, Q150A, Q150D, Q150E, Q150F, Q150G, Q150H,Q150I, Q150K, Q150L, Q150M, Q150N, Q150P, Q150R, Q150S, Q150T, Q150V,Q150W, Q150Y, F151*, F151A, F151D, F151E, F151G, F151H, F151I, F151K,F151L, F151M, F151N, F151P, F151Q, F151R, F151S, F151T, F151V, F151W,F151Y, D152*, D152A, D152E, D152F, D152G, D152H, D152I, D152K, D152L,D152M, D152N, D152P, D152Q, D152R, D152S, D152T, D152V, D152W, D152Y,P153*, P153A, P153D, P153E, P153F, P153G, P153H, P153I, P153K, P153L,P153M, P153N, P153Q, P153R, P153S, P153T, P153V, P153W, P153Y, S154*,S154A, S154D, S154E, S154F, S154G, S154H, S154I, S154K, S154L, S154M,S154N, S154P, S154Q, S154R, S154T, S154V, S154W, S154Y, K155*, K155A,K155D, K155E, K155F, K155G, K155H, K155I, K155L, K155M, K155N, K155P,K155Q, K155R, K155S, K155T, K155V, K155W, K155Y, P156*, P156A, P156D,P156E, P156F, P156G, P156H, P156I, P156K, P156L, P156M, P156N, P156Q,P156R, P156S, P156T, P156V, P156W, P156Y, Q157*, Q157A, Q157D, Q157E,Q157F, Q157G, Q157H, Q157I, Q157K, Q157L, Q157M, Q157N, Q157P, Q157R,Q157S, Q157T, Q157V, Q157W, Q157Y, Q158*, Q158A, Q158D, Q158E, Q158F,Q158G, Q158H, Q158I, Q158K, Q158L, Q158M, Q158N, Q158P, Q158R, Q158S,Q158T, Q158V, Q158W, Q158Y, T159*, T159A, T159D, T159E, T159F, T159G,T159H, T159I, T159K, T159L, T159M, T159N, T159P, T159Q, T159R, T159S,T159V, T159W, T159Y, K160*, K160A, K160D, K160E, K160F, K160G, K160H,K160I, K160L, K160M, K160N, K160P, K160Q, K160R, K160S, K160T, K160V,K160W, K160Y, G161*, G161A, G161D, G161E, G161F, G161H, G161I, G161K,G161L, G161M, G161N, G161P, G161Q, G161R, G161S, G161T, G161V, G161W,G161Y, T162A, T162D, T162E, T162F, T162G, T162H, T162I, T162K, T162L,T162M, T162N, T162P, T162Q, T162R, T162S, T162T, T162V, T162W, T162Y,W163*, W163A, W163D, W163E, W163F, W163G, W163H, W163I, W163K, W163L,W163M, W163N, W163P, W163Q, W163R, W163S, W163T, W163V, W163Y, F164*,F164A, F164D, F164E, F164G, F164H, F164I, F164K, F164L, F164M, F164N,F164P, F164Q, F164R, F164S, F164T, F164V, F164W, F164Y, Q165*, Q165A,Q165D, Q165E, Q165F, Q165G, Q165H, Q165I, Q165K, Q165L, Q165M, Q165N,Q165P, Q165R, Q165S, Q165T, Q165V, Q165W, Q165Y, I166*, I166A, I166D,I166E, I166F, I166G, I166H, I166K, I166L, I166M, I166N, I166P, I166Q,I166R, I166S, I166T, I166V, I166W, I166Y, T167*, T167A, T167D, T167E,T167F, T167G, T167H, T167I, T167K, T167L, T167M, T167N, T167P, T167Q,T167R, T167S, T167V, T167W, T167Y, K168*, K168A, K168D, K168E, K168F,K168G, K168H, K168I, K168L, K168M, K168N, K168P, K168Q, K168R, K168S,K168T, K168V, K168W, K168Y, F169*, F169A, F169D, F169E, F169G, F169H,F169I, F169K, F169L, F169M, F169N, F169P, F169Q, F169R, F169S, F169T,F169V, F169W, F169Y, T170*, T170A, T170D, T170E, T170F, T170G, T170H,T170I, T170K, T170L, T170M, T170N, T170P, T170Q, T170R, T170S, T170V,T170W, T170Y, G171*, G171A, G171D, G171E, G171F, G171H, G171I, G171K,G171L, G171M, G171N, G171P, G171Q, G171R, G171S, G171T, G171V, G171W,G171Y, A172*, A172D, A172E, A172F, A172G, A172H, A172I, A172K, A172L,A172M, A172N, A172P, A172Q, A172R, A172S, A172T, A172V, A172W, A172Y,A173*, A173D, A173E, A173F, A173G, A173H, A173I, A173K, A173L, A173M,A173N, A173P, A173Q, A173R, A173S, A173T, A173V, A173W, A173Y, G174*,G174A, G174A, G174D, G174DG174A, G174D, G174E, G174E, G174F, G174FG174E,G174F, G174H, G174H, G174I, G1741G174H, G174I, G174K, G174K, G174L,G174LG174K, G174L, G174M, G174M, G174N, G174N, G174M, G174N, G174P,G174P, G174Q, G174QG174P, G174Q, G174R, G174R, G174S, G174SG174R, G174S,G174T, G174T, G174V, G174VG174T, G174V, G174W, G174W, G174Y, G174YG174W,G174Y, P175*, P175A, P175D, P175E, P175F, P175G, P175H, P175I, P175K,P175L, P175M, P175N, P175Q, P175R, P175S, P175T, P175V, P175W, P175Y,Y176*, Y176A, Y176D, Y176E, Y176F, Y176G, Y176H, Y176I, Y176K, Y176L,Y176M, Y176N, Y176P, Y176Q, Y176R, Y176S, Y176T, Y176V, Y176W, K178*,K178A, K178D, K178E, K178F, K178G, K178H, K178I, K178L, K178M, K178N,K178P, K178Q, K178R, K178S, K178T, K178V, K178W, K178Y, A179*, A179D,A179E, A179F, A179G, A179H, A179I, A179K, A179L, A179M, A179N, A179P,A179Q, A179R, A179S, A179T, A179V, A179W, A179Y, L180*, L180A, L180D,L180E, L180F, L180G, L180H, L180I, L180K, L180M, L180N, L180P, L180Q,L180R, L180S, L180T, L180V, L180W, L180Y, G181*, G181A, G181D, G181E,G181F, G181H, G181I, G181K, G181L, G181M, G181N, G181P, G181Q, G181R,G181S, G181T, G181V, G181W, G181Y, S182*, S182A, S182D, S182E, S182F,S182G, S182H, S182I, S182K, S182L, S182M, S182N, S182P, S182Q, S182R,S182T, S182V, S182W, S182Y, N183*, N183A, N183D, N183E, N183F, N183G,N183H, N183I, N183K, N183L, N183M, N183P, N183Q, N183R, N183S, N183T,N183V, N183W, N183Y, D184*, D184A, D184E, D184F, D184G, D184H, D184I,D184K, D184L, D184M, D184N, D184P, D184Q, D184R, D184S, D184T, D184V,D184W, D184Y, K185*, K185A, K185D, K185E, K185F, K185G, K185H, K185I,K185L, K185M, K185N, K185P, K185Q, K185R, K185S, K185T, K185V, K185W,K185Y, S186*, S186A, S186D, S186E, S186F, S186G, S186H, S186I, S186K,S186L, S186M, S186N, S186P, S186Q, S186R, S186T, S186V, S186W, S186Y,V187*, V187A, V187D, V187E, V187F, V187G, V187H, V187I, V187K, V187L,V187M, V187N, V187P, V187Q, V187R, V187S, V187T, V187W, V187Y, D189*,D189A, D189E, D189F, D189G, D189H, D189I, D189K, D189L, D189M, D189N,D189P, D189Q, D189R, D189S, D189T, D189V, D189W, D189Y, K190*, K190A,K190D, K190E, K190F, K190G, K190H, K190I, K190L, K190M, K190N, K190P,K190Q, K190R, K190S, K190T, K190V, K190W, K190Y, N191*, N191A, N191D,N191E, N191F, N191G, N191H, N191I, N191K, N191L, N191M, N191P, N191Q,N191R, N191S, N191T, N191V, N191W, N191Y, K192*, K192A, K192D, K192E,K192F, K192G, K192H, K192I, K192L, K192M, K192N, K192P, K192Q, K192R,K192S, K192T, K192V, K192W, K192Y, N193*, N193A, N193D, N193E, N193F,N193G, N193H, N193I, N193K, N193L, N193M, N193P, N193Q, N193R, N193S,N193T, N193V, N193W, N193Y, I194*, I194A, I194D, I194E, I194F, I194G,I194H, I194K, I194L, I194M, I194N, I194P, I194Q, I194R, I194S, I194T,I194V, I194W, I194Y, A195*, A195D, A195E, A195F, A195G, A195H, A195I,A195K, A195L, A195M, A195N, A195P, A195Q, A195R, A195S, A195T, A195V,A195W, A195Y, G196*, G196A, G196D, G196E, G196F, G196H, G196I, G196K,G196L, G196M, G196N, G196P, G196Q, G196R, G196S, G196T, G196V, G196W,G196Y, D197*, D197A, D197E, D197F, D197G, D197H, D197I, D197K, D197L,D197M, D197N, D197P, D197Q, D197R, D197S, D197T, D197V, D197W, D197Y,W198*, W198A, W198D, W198E, W198F, W198G, W198H, W198I, W198K, W198L,W198M, W198N, W198P, W198Q, W198R, W198S, W198T, W198V, W198Y, G199*,G199A, G199D, G199E, G199F, G199H, G199I, G199K, G199L, G199M, G199N,G199P, G199Q, G199R, G199S, G199T, G199V, G199W, G199Y, F200*, F200A,F200D, F200E, F200G, F200H, F200I, F200K, F200L, F200M, F200N, F200P,F200Q, F200R, F200S, F200T, F200V, F200W, F200Y, D201*, D201A, D201E,D201F, D201G, D201H, D201I, D201K, D201L, D201M, D201N, D201P, D201Q,D201R, D201S, D201T, D201V, D201W, D201Y, P202*, P202A, P202D, P202E,P202F, P202G, P202H, P202I, P202K, P202L, P202M, P202N, P202Q, P202R,P202S, P202T, P202V, P202W, P202Y, A203*, A203D, A203E, A203F, A203G,A203H, A203I, A203K, A203L, A203M, A203N, A203P, A203Q, A203R, A203S,A203T, A203V, A203W, A203Y, K204*, K204A, K204D, K204E, K204F, K204G,K204H, K204I, K204L, K204M, K204N, K204P, K204Q, K204R, K204S, K204T,K204V, K204W, K204Y, W205*, W205A, W205D, W205E, W205F, W205G, W205H,W205I, W205K, W205L, W205M, W205N, W205P, W205Q, W205R, W205S, W205T,W205V, W205Y, A206*, A206D, A206E, A206F, A206G, A206H, A206I, A206K,A206L, A206M, A206N, A206P, A206Q, A206R, A206S, A206T, A206V, A206W,A206Y, Y207*, Y207A, Y207D, Y207E, Y207F, Y207G, Y207H, Y207I, Y207K,Y207L, Y207M, Y207N, Y207P, Y207Q, Y207R, Y207S, Y207T, Y207V, Y207W,Q208*, Q208A, Q208D, Q208E, Q208F, Q208G, Q208H, Q208I, Q208K, Q208L,Q208M, Q208N, Q208P, Q208R, Q208S, Q208T, Q208V, Q208W, Q208Y, Y209*,Y209A, Y209D, Y209E, Y209F, Y209G, Y209H, Y209I, Y209K, Y209L, Y209M,Y209N, Y209P, Y209Q, Y209R, Y209S, Y209T, Y209V, Y209W, D210*, D210A,D210E, D210F, D210G, D210H, D210I, D210K, D210L, D210M, D210N, D210P,D210Q, D210R, D210S, D210T, D210V, D210W, D210Y, E211*, E211A, E211D,E211F, E211G, E211H, E211I, E211K, E211L, E211M, E211N, E211P, E211Q,E211R, E211S, E211T, E211V, E211W, E211Y, K212*, K212A, K212D, K212E,K212F, K212G, K212H, K212I, K212L, K212M, K212N, K212P, K212Q, K212R,K212S, K212T, K212V, K212W, K212Y, N213*, N213A, N213D, N213E, N213F,N213G, N213H, N213I, N213K, N213L, N213M, N213P, N213Q, N213R, N213S,N213T, N213V, N213W, N213Y, N214*, N214A, N214D, N214E, N214F, N214G,N214H, N214I, N214K, N214L, N214M, N214P, N214Q, N214R, N214S, N214T,N214V, N214W, N214Y, K215*, K215A, K215D, K215E, K215F, K215G, K215H,K215I, K215L, K215M, K215N, K215P, K215Q, K215R, K215S, K215T, K215V,K215W, K215Y, F216*, F216A, F216D, F216E, F216G, F216H, F216I, F216K,F216L, F216M, F216N, F216P, F216Q, F216R, F216S, F216T, F216V, F216W,F216Y, N217*, N217A, N217D, N217E, N217F, N217G, N217H, N217I, N217K,N217L, N217M, N217P, N217Q, N217R, N217S, N217T, N217V, N217W, N217Y,Y218*, Y218A, Y218D, Y218E, Y218F, Y218G, Y218H, Y218I, Y218K, Y218L,Y218M, Y218N, Y218P, Y218Q, Y218R, Y218S, Y218T, Y218V, Y218W, V219*,V219A, V219D, V219E, V219F, V219G, V219H, V219I, V219K, V219L, V219M,V219N, V219P, V219Q, V219R, V219S, V219T, V219W, V219Y, G220*, G220A,G220D, G220E, G220F, G220H, G220I, G220K, G220L, G220M, G220N, G220P,G220Q, G220R, G220S, G220T, G220V, G220W, G220Y, K221*, K221A, K221D,K221E, K221F, K221G, K221H, K221I, K221L, K221M, K221N, K221P, K221Q,K221R, K221S, K221T, K221V, K221W and K221Y compared to SEQ ID NO: 1.14. The method according to any of embodiments 11-13, wherein the DNasevariant has at least 60%, such as at least 70%, at least 80%, at least90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least99%, sequence identity to the mature polypeptide of SEQ ID NO: 1.Assays and Detergent CompositionsComposition of Model Detergent A (Liquid)

Ingredients: 12% LAS, 11% AEO Biosoft N25-7 (NI), 7% AEOS (SLES), 6% MPG(monopropylene glycol), 3% ethanol, 3% TEA, 2.75% cocoa soap, 2.75% soyasoap, 2% glycerol, 2% sodium hydroxide, 2% sodium citrate, 1% sodiumformiate, 0.2% DTMPA and 0.2% PCA (all percentages are w/w)

Composition of Model Detergent T (Powder)

Ingredients: 11% LAS, 2% AS/AEOS, 2% soap, 3% AEO, 15.15% sodiumcarbonate, 3% sodium silicate, 18.75% zeolite, 0.15% chelant, 2% sodiumcitrate, 1.65% AA/MA copolymer, 2.5% CMC and 0.5% SRP (all percentagesare w/w).

Composition of Model Detergent X (Powder)

Ingredients: 16.5% LAS, 15% zeolite, 12% sodium disilicate, 20% sodiumcarbonate, 1% sokalan, 35.5% sodium sulfate (all percentages are w/w).

Assay I: Testing of DNase Activity

DNase activity was determined on DNase Test Agar with Methyl Green (BD,Franklin Lakes, N.J., USA), which was prepared according to the manualfrom supplier. Briefly, 21 g of agar was dissolved in 500 ml water andthen autoclaved for 15 min at 121° C. Autoclaved agar was temperated to48° C. in water bath, and 20 ml of agar was poured into petridishes withand allowed to solidify by incubation o/n at room temperature. Onsolidified agar plates, 5 μl of enzyme solutions are added and DNaseactivity is observed as colorless zones around the spotted enzymesolutions.

Assay II: Analysis of E-2-Nonenal on Textile Using an Electronic Nose.

One way of testing for the presence of malodor on textiles is by usingE-2-Nonenal as a marker for the malodor, as this compound contributes tothe malodor on laundry.

Add a solution of E-2-nonenal to a 5 cm×5 cm textile swatch and placethe swatch in a 20 mL glass vial for GC analysis and cap the vial.Analyze 5 mL headspace from the capped vials in a Heracles II Electronicnose from Alpha M.O.S., France (double column gas chromatograph with 2FIDs, column 1: MXT5 and column 2: MXT1701) after 20 minutes incubationat 40° C.

Methods

General methods of PCR, cloning, ligation nucleotides etc. arewell-known to a person skilled in the art and may for example be foundin in “Molecular cloning: A laboratory manual”, Sambrook et al. (1989),Cold Spring Harbor lab., Cold Spring Harbor, N.Y.; Ausubel, F. M. et al.(eds.); “Current protocols in Molecular Biology”, John Wiley and Sons,(1995); Harwood, C. R., and Cutting, S. M. (eds.); “DNA Cloning: APractical Approach, Volumes I and II”, D. N. Glover ed. (1985);“Oligonucleotide Synthesis”, M. J. Gait ed. (1984); “Nucleic AcidHybridization”, B. D. Hames & S. J. Higgins eds (1985); “A PracticalGuide To Molecular Cloning”, B. Perbal, (1984).

EXAMPLES Example 1: Preparation of DNase Variants

Site-directed variants (SD) were generated using specific primers thatcontained the desired new mutation. The new codon selected was the codonwith the highest natural abundance for the specific amino acid inAspergillus oryzae. The transformation substrate was made in two roundsof PCR:

1) Separate amplification of N-terminal and C-terminal fragment,relative to the mutation site. The N-terminal fragment was amplifiedusing a universal forward primer and a position specific reverse primer.The C-terminal fragment was amplified using a mutation specific forwardprimer and universal reverse primer. Both universal primers arecomplementary to sequences necessary for homologous integration inAspergillus genome.2) Assembly of the N-terminal and C-terminal fragments by fusion (SOE)PCR.

The resulting transformation substrate was transformed into Aspergillusoryzae and transformants selected for by growth on nitrate as solenitrogen source. Three single colonies of each type were picked into MTPand grown for 4 days at 30° C. in broth specific for Aspergillus. Thesupernatants were used for screening.

Example 2: Testing DNase Variants for Stability

The stability of DNase variants compared to DNase with SEQ ID NO: 1 wasdetermined by incubating the DNase samples under defined conditions(“stress conditions”) in a model detergent solution (Model detergent A).The variants were tested using two different set of conditions stressconditions given by temperature and duration.

A) Set A was chosen such that the remaining activity of the DNase withSEQ ID NO: 1 after the incubation is equal to approximately 50% of theactivity of a similar sample incubated under defined conditions(“reference conditions”) that do not lead to loss of activity whenincubated for the same duration. The samples were normalized by activityas described below and the normalized samples stressed in PCR machinesat 50° C. for 20 hrs. The activity after incubation under stressconditions or reference conditions was determined using viscosity assaydescribed below.

B) Set B was chosen such that the remaining activity of the DNase withSEQ ID NO: 1 after the incubation is equal to approximately 20% of theactivity of a similar sample incubated under defined conditions(“reference conditions”) that do not lead to loss of activity whenincubated for the same duration. The samples were normalized by ELISA asdescribed below and the normalized samples stressed in heating cabinetat 50° C. for 72 hrs. The activity after incubation under stressconditions or reference conditions was determined using viscosity assaydescribed below.

Normalization of Samples by Activity

A 96 well plate with supernatants of Aspergillus cultures expressingDNase variants was shaken for 2 minutes at 600 rpm on an BioSanThermo-Shaker (PST-100HL) before loaded on the Hamilton STAR robotequipped with a 96 channel TADM head which allows the simultaneousmeasuring of pressure in the head space of each single pipette channel.10 μl of the supernatant from each of the 96 wells is transferred to 96wells on an empty plate with each well containing 90 μl 50 mM Tris-HCl,pH 7. From each of the 96 wells 5 μl of this 10× dilution is transferredto a new plate with each well containing 195 μl of DNA substrate (3.3 mgDNA/ml, 50 mM Tris-HCl, 5 mM MgCl2 at pH7; DNA: Deoxyribonucleic acidsodium salt from salmon testes, Sigma D1626).

The Hamilton STAR measures the viscosity using ViPr assay technology asdescribed in (WO2011/107472 A1) of the solution in all 96 wells for 30min at a 1 min interval by aspirating 100 μl with a standard clear CORE300 μl tip at a speed of 50 μl/s with the addition of the diluted DNasesample after the second measurement. The solution is dispensed back tothe original well at 10 μl/s.

The pressure data during the aspiration step from all of the 96 wellsare collected and the pressure value at 1000 ms after start of theaspiration is taken for calculation of the DNase activity in Relativeunits (RU) from a standard that is loaded on each plate. The standard oneach plate is 1, 5 and 10 ppm wt DNase and is in replica. Relative unitsare calculated by division of the buffer value (blank; 1000 ms) by thepressure value (1000 ms) of a given sample subtracted by 1. Relativeunits for the standard curve are plotted against the concentration and alinear curve fitting is applied. From the RU values of the samples theconcentration in ppm can be obtained using the standard curve.

Based on the concentration determination the original samples arenormalized to 10 ppm in 50 mM Tris HCL pH7 or the maximum obtainableconcentration in case of low expressing variants for the subsequentstability screening.

Normalization of Samples by ELISA Assay for Protein ContentDetermination

The DNase variants and the concentration of the reference DNase (SEQ IDNO: 1) was alternative determined by ELISA after a 9750-fold dilution ofsupernatants in TBS-T buffer (20 mM Tris, 150 mM NaCl, 0.1 v/v Tween 20,pH 7.5)). For protein quantification the sandwich ELISA technique wasused. Microplate wells coated with rabbit polyclonal anti-proteinantibodies constitute the solid phase. Test samples, standards, andcontrols are added to the coated wells and incubated with incubationbuffer. If the protein of interest is present in the test sample, itwill be captured and immobilised by the protein-specific antibody coatedon the wells. After incubation and washing, another polyclonalanti-protein antibody is added to the wells. This antibody is labeledwith horseradish peroxidase (HRP). The HRP-labeled antibody binds to theimmobilised protein of interest, thus forming a sandwich. Afterincubation and washing, an enzyme substrate is added to the wells andincubated. The degree of enzymatic activity of the immobilised HRP isdetermined by measuring the optical density of the oxidized enzymaticproduct in the wells at 620 nm. The absorbance at 620 nm is proportionalto the amount of protein in the test sample. A set of standards is usedto generate a standard curve of absorbance versus protein concentration.This curve is used for calculating the concentrations of test samplesand controls. Based on the concentration determination the originalsamples are normalized to 10 ppm in 50 mM Tris HCL pH7.

Stability Assay

A DNase standard (0, 5 and 10 ppm) is added to the Normalization platecontaining the samples normalized to 10+ ppm or the maximum obtainableconcentration when below 10 ppm. 12 μl sample from the normalized plateis added to 228 μl Model detergent A in a well of a standard 96 wellmicrotiter plate. Detergent and samples are mixed by shaking for 90seconds at 2000 rpm using a Hamilton Heater Shaker (HHS) that is mountedon the Hamilton STAR. After the mixing 200 μl of the sample is aspiratedand 80 μl of the sample is added to all wells of two 96 well PCR plates.

The plates are sealed and one plate is stored at 4° C. (unstressed)while the replica plate is incubated at 50° C. (stressed) in a PCRmachine for 20 hours or in a heating cabinet for 72 hrs respectively.

Residual Activity Determination

10 μl of sample from column 1-6 or 7-12 from the stress and unstressedplate respectively are transferred to a standard 96 well plate with eachwell filled with 190 μl of DNase substrate (3.3 mg DNA/ml, 50 mMTris-HCl, 5 mM MgCl2 at pH7; DNA: Deoxyribonucleic acid sodium salt fromsalmon testes, Sigma D1626). The stressed and unstressed samples aresubsequently mixed by 5 consecutive aspirations and dispense steps witha volume of 100 μl. After mixing the plates are incubated for 1.5 h at37° C. followed by 30 min incubation at room temperature. Subsequentlythe viscosity of the samples was measured by 5 consecutive aspirationsand dispenses steps with a volume of 100 μl and an aspirate speed of 50μl/s a Hamilton STAR equipped with a 96 channel TADM head using VIPR(WO2011/107472 A1). The pressure data during the aspiration step fromall of the 96 wells are collected and the average pressure value at 1000ms after start of the aspiration is calculated for the for the 4 lastaspiration steps; both for the stressed and unstressed samples includingthe buffer blanks and the reference samples (SEQ ID NO: 1) located oneach plate. Relative units are calculated by division of the buffervalue (blank; 1000 ms) by the pressure value (1000 ms) of a given samplesubtracted by 1. This is calculated for both the stressed and unstressedsample. The ratio between RU for the stressed and unstressed sample iscalculated in percent which is equivalent to the residual activity (RA%).

Half-Life Improvement Factor

The half-life improvement factor (IF) correlates the residual activitypercentage (RA %) of a DNase variant with that of the reference DNase(SEQ ID NO: 1). Calculation of the improvement factor based on residualactivity percentage was done according to the following equation:IF=(ln(0.5)/ln(RA % of variant))/(ln(0.5)/ln(RA % of SEQ ID NO: 1))The value for RA % of reference DNase (SEQ ID NO: 1) employed was alwayscalculated from samples residing on the same screening plate as theDNase variant. The following criteria was used for qualification of datafor a given sample: The pressure value (1000 ms) for the unstressedsample must be lower than −400 Pa, the calculated relative unit for theunstressed sample must be between 0.2 and 2, the calculated residualactivity percentage (RA %) should be between 0 and 100% and RA % ofreference DNase must be between 10 and 60%.

An improvement factor that is greater than 1 (IF>1) indicates animproved stability of a variant as compared to the reference, while anIF of 1 (IF=1) identifies a variant which is on par with the reference,and an IF of less than 1 (IF<1) identifies a variant that is less stablethe reference.

The improvement factor was ranked on a scale from 1-3 using a scale thatconsidered the variation in the calculated improvement factor betweenthree biological replicates. Also, two different scales were employed toaccount for IF variations between set A and set B when changingscreening conditions. The intervals used for ranking IF from set A were:

1: IF<1.25

2: 1.25<=IF<2

3: 2<=IF

The intervals used for ranking IF from set B were:

1: IF<1.5

2: 1.5<=IF<3

3: 3<=IF

For each variant, the raking from three biological replicates wascombined to an overall score. The median of the ranking was used whenthe value could be calculated for all three biological replicates. Thelowest ranking was used when two biological replicates were represented,while the ranking was disregarded when only a singular value wasobtainable.

The assay rankings and scores for a selection of DNase variants arelisted in the table 1 below:

TABLE 1 Rank Rank Position Variant (set A) (set B) 4 N4E 2 17 L17E 2 19T19A 2 19 T19G 2 19 T19I 2 36 K36P 2 38 Q38P 2 39 S39V 2 39 S39R 3 40A40P 2 40 A40H 2 41 L41T 3 41 L41H 2 45 V45H 2 51 L51G 2 53 K53T 2 53K53P 2 54 G54P 2 55 A55P 2 56 P56* 2 57 N57H 2 64 E64A 2 64 E64Q 2 64E64R 2 64 E64T 2 64 E64I 3 64 E64S 3 66 T66H 2 67 K67A 2 67 K67T 2 68N68V 2 68 N68P 3 68 N68I 2 68 N68H 2 69 S69A 2 69 S69D 2 69 S69E 2 69S69K 2 69 S69L 2 69 S69W 2 69 S69Y 2 69 S69Q 3 70 N70T 3 70 N70H 2 70N70G 2 71 R71T 3 72 D72E 2 74 S74H 2 74 S74G 2 75 G75I 2 77 N77T 2 82K82P 2 82 K82I 2 83 D83T 2 83 D83P 3 83 D83I 2 83 D83H 2 83 D83G 2 84P84H 2 84 P84* 2 85 Q85T 2 85 Q85P 2 85 Q85H 2 86 K86T 2 86 K86P 2 86K86H 2 88 G88P 2 88 G88H 2 88 G88* 2 91 A91P 2 99 W99T 3 101 A101W 2 105K105E 2 105 K105N 2 105 K105T 2 105 K105D 3 106 S106T 2 115 S115T 2 116L116I 2 135 Q135L 3 136 G136L 3 138 V138I 2 138 V138L 2 138 V138P 2 138V138Q 2 139 L139A 2 139 L139* 3 140 N140R 2 140 N140L 3 140 N140A 2 141G141L 2 151 F151R 2 152 D152Y 2 152 D152L 2 152 D152I 2 152 D152A 2 153P153E 2 154 S154R 2 162 T162R 2 163 W163E 2 164 F164R 2 166 I166Y 2 166I166R 2 168 K168N 2 169 F169R 2 169 F169E 2 173 A173I 2 173 A173R 2 173A173T 2 179 A179* 2 180 L180* 2 182 S182R 2 183 N183E 2 184 D184I 2 185K185Y 2 186 S186I 2 189 D189G 2 189 D189H 2 212 K212G 2 212 K212P 2 215K215I 2

What is claimed is:
 1. A DNase variant, which: (a) has at least 90%sequence identity to SEQ ID NO: 1; (b) comprises a substitution at oneor more positions corresponding to the positions selected from the groupconsisting of 32, 35, 105, 111 and 181 of SEQ ID NO: 1; (c) has DNaseactivity; and (d) has improved stability as compared to the polypeptideof SEQ ID NO:
 1. 2. The DNase variant of claim 1, wherein the varianthas at least 95% sequence identity to the polypeptide of SEQ ID NO: 1.3. The DNase variant of claim 1, which comprises a substitution at aposition corresponding to position 32 of SEQ ID NO: 1 with Ala, Glu,Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val,Trp or Tyr.
 4. The DNase variant of claim 1, which comprises asubstitution at a position corresponding to position 35 of SEQ ID NO: 1with Ala, Asp, Glu, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln,Arg, Ser, Thr, Trp or Tyr.
 5. The DNase variant of claim 1, whichcomprises a substitution at a position corresponding to position 105 ofSEQ ID NO: 1 with Ala, Asp, Glu, Phe, Gly, His, Ile, Leu, Met, Asn, Gln,Pro, Arg, Ser, Thr, Val, Trp or Tyr.
 6. The DNase variant of claim 5,wherein the substitution at a position corresponding to position 105 ofSEQ ID NO: 1 is K105E, K105N, K105T or K105D.
 7. The DNase variant ofclaim 2, which comprises a substitution at a position corresponding toposition 105 of SEQ ID NO: 1 with Ala, Asp, Glu, Phe, Gly, His, Ile,Leu, Met, Asn, Gln, Pro, Arg, Ser, Thr, Val, Trp or Tyr.
 8. The DNasevariant of claim 7, wherein the substitution at a position correspondingto position 105 of SEQ ID NO: 1 is K105E, K105N, K105T or K105D.
 9. TheDNase variant of claim 1, which comprises a substitution at a positioncorresponding to position 111 of SEQ ID NO: 1 with Asp, Glu, Phe, Gly,His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val, Trp or Tyr.10. The DNase variant of claim 1, which comprises a substitution at aposition corresponding to position 181 of SEQ ID NO: 1 with Ala, Asp,Glu, Phe, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Thr, Val,Trp or Tyr.
 11. The DNase variant of claim 1, which is a variant of aparent polypeptide wherein the parent polypeptide is the polypeptide ofSEQ ID NO:
 1. 12. The DNase variant of claim 1, wherein the total numberof amino acid substitutions compared to SEQ ID NO: 1 is between 1-20.13. The DNase variant of claim 1, further comprising one or moreadditional substitutions at one or more positions corresponding topositions 4, 17, 19, 36, 38, 39, 40, 41, 45, 51, 53, 54, 55, 57, 64, 66,67, 68, 69, 70, 71, 72, 74, 75, 77, 82, 83, 84, 85, 86, 88, 91, 99, 101,106, 115, 116, 135, 136, 138, 139, 140, 141, 151, 152, 153, 154, 162,163, 164, 166, 168, 169, 173, 182, 183, 184, 185, 186, 189, 212 and 215of SEQ ID NO:
 1. 14. The DNase variant of claim 13, wherein the one ormore additional substitutions are selected from the group consisting ofN4E, L17E, T19A, T19G, T19I, K36P, Q38P, S39V, S39R, A40P, A4OH, L41T,L41H, V45H, L51G, K53T, K53P, G54P, A55P, N57H, E64A, E64Q, E64R, E64T,E64I, E64S, T66H, K67A, K67T, N68V, N68P, N68I, N68H, S69A, S69D, S69E,S69K, S69L, S69W, S69Y, S69Q, N70T, N70H, N70G, R71T, D72E, S74H, 574G,G75I, N77T, K82P, K82I, D83T, D83P, D83I, D83H, D83G, P84H, Q85T, Q85P,Q85H, K86T, K86P, K86H, G88P, G88H, A91P, W99T, A101W, S106T, S115T,L116I, Q135L, G136L, V138I, V138L, V138P, V138Q, L139A, N140R, N140L,N140A, G141L, F151R, D152Y, D152L, D152I, D152A, P153E, S154R, T162R,W163E, F164R, I166Y, I166R, K168N, F169R, F169E, A173I, A173R, A173T,S182R, N183E, D184I, K185Y, S186I, D189G, D189H, K212G, K212P and K215I.15. The DNase variant of claim 1, wherein the DNase variant has anImprovement Factor (IF) of at least 1.1 as compared to the polypeptideshown in SEQ ID NO:
 1. 16. A composition comprising: (a) at least 0.002ppm of the DNase variant of claim 1; (b) 2 wt % to 60 wt % of at leastone surfactant; and (c) 5 wt % to 50 wt % of at least one builder. 17.The composition of claim 16, wherein the composition is a detergentcomposition.
 18. The composition of claim 16, further comprising one ormore additional enzymes selected from the group consisting of amylases,catalases cellulases, haloperoxygenases, lipases, mannanases,xyloglucanases, pectinases, pectin lyases, peroxidases, proteases,xanthanases, and any mixture thereof.
 19. The composition of claim 16,which is in the form of a bar, a homogenous tablet, a tablet having twoor more layers, a pouch having one or more compartments, a regular orcompact powder, a granule, a paste, a gel, or a regular, compact orconcentrated liquid.
 20. A method of cleaning laundry or a hard surface,comprising washing the laundry or the hard surface with the compositionof claim 16.